Vitamin d analogues, compositions comprising said analogues and their use

ABSTRACT

Compounds according to formula I 
                         
in which formula R1 and R2, which may be the same or different, represent halogen, (C 1 -C 6 )hydrocarbyl, optionally substituted with one or two hydroxyl group or one or more fluorine atoms, or, together with the carbon atom to which they are both attached, R1 and R2 form a (C 3 -C 6 )carbocyclic ring, or one of R1 and R2 taken together with R3 forms a direct bond, such that a triple bond is constituted, or R1 and R2 represent both hydrogen; R3 when not forming a direct bond with one of R1 and R2 represents hydrogen or (C 1 -C 3 )hydrocarbyl; X represents (E)-ethylene, (Z)-ethylene, ethynylene, or a bond; Y and Z independently represent hydrogen or methyl; and prodrugs and stereo isomeric forms thereof are provided together with their use in therapy, and their use in the manufacture of medicaments.

This National Phase PCT application claims priority under 35 U.S.C.119(e) on U.S. Provisional Application No(s). 60/420,783 filed on Oct.24, 2002 and under 35 U.S.C. 119(a) on Patent Application No(s). PA 200201608 filed in Japan on Oct. 23, 2002, all of which which are herebyincorporated by reference.

FIELD OF INVENTION

This invention relates to novel vitamin D analogues, to their use intherapy, to pharmaceutical compositions comprising said analogues, tomethods of treatment comprising the administration of said analogues topatients in need thereof, and to the use of said analogues in themanufacture of medicaments.

BACKGROUND OF THE INVENTION

Over the last decades there has been a growing understanding of thebiological effects of vitamin D. The classical actions of vitamin Dinvolve calcium and phosphate absorption from the intestines, which isvital to the mineral balance and to the build-up and maintenance ofbones. Another primary action of vitamin D is the regulation of theexcretion of the parathyroid hormone (PTH) from the parathyroid glands.Vitamin D inhibits the production of the parathyroid hormone, so that alow level of vitamin D in the blood will lead to a high levelparathyroid hormone, and vice versa. Vitamin D exerts its effect throughan intriguing mechanism whereby the production of the mRNA which istranslated into the parathyroid hormone, or a proform thereof, isinhibited. The impact of vitamin D in biological systems, however,reaches beyond these effects. Vitamin D appears to have profound effectson muscles, the immune system, the reproductive system, and cellproliferation and differentiation. Cells holding the vitamin D receptor(VDR) have, in fact, been found in many parts of the body, including theintestines, kidneys, prostate, bone, bone marrow, parathyroid glands,skin, liver, muscle and lymphoid tissue. The widespread existence of VDRhave made vitamin D and analogues thereof attractive compounds for thetreatment of various diseases including cancer, skin and bone diseasesand autoimmune diseases.

The invention relates to a novel class of vitamin D analogues that showa potent suppressive effect on the secretion of parathyroid hormone,i.e. which can be used in the treatment of secondary hyperparathyroidism(s-HPT). A crucial structural element in active vitamin D are the twohydroxyl groups in positions 1 and 25. In contrast to that, thecompounds of the present invention are characterized by a blocking ofthe 25-position, so that they do not have hydroxyl groups in thatposition, nor can they be hydroxylated in that position in vivo by aP450-like enzyme.

Vitamin D analogues with some structural resemblance to the compounds ofthe present invention have previously been disclosed. As an example,WO95/02577 teaches compounds of the formula

WO91/00855 discloses compounds of the formula

and Onisko, Tetrahedron Lett., 1107-1108, 13, 1977 discloses a compoundof the formula

which is useful for inhibition of liver enzymes responsible forhydroxylation of vitamin D₃ to 25-OH vitamin D₃.

Finally, Bogoslovsky et al, Vitamin D—Basic Research and its ClinicalApplication, proceedings of the Fourth Workshop on Vitamin D, BerlinWest Germany 1979, A. W. Norman et al (Eds.), p 1257-1259, Walter deGruyter, Berlin 1979, discloses a synthetic study including thepreparation of3(S)-hydroxy-9,10-secocholesta-5(Z),7(E),10(19),22(E),24-penta-ene. Thisreference, however, does not disclose any biological data on thisparticular compound.

Vitamin D and analogues thereof are already used in the treatment ofs-HPT. Paricalcitol (19-nor-1,25-dihydroxy-vitamin D₂) anddoxercalciferol (1α-hydroxy-vitamin D₂) are approved in the USA fortreatment of s-HPT, and 22-oxa-calcitriol (maxacalcitol) andhexafluoro-calcitriol (falecalcitriol) are approved in Japan [Malluche,Kidney Int., 367-374, 62, 2002]. Moreover, calcitriol itself and aprodrug thereof 1α(OH)D₃ are also used in the treatment and prophylaxisof s-HPT [Brandi, Nephrol Dial Transplant, 829-842, 17, 2002].

All therapeutic interventions which include administration of vitamin Dand analogues thereof must pay attention to the adverse side effectsoften associated with this kind of therapy, in particular the calcemiceffects of vitamin D compounds. These side effects may severely restrictor even prevent the use of such compounds, in spite of other clinicallypositive effects. The present invention therefore seeks to providevitamin D analogues which have a reduced calcemic effect while retaininga suppressive effect on the secretion of the parathyroid hormone.

SUMMARY OF THE INVENTION

Accordingly, the present invention provides compounds represented byformula I

in which formula

-   R1 and R2, which may be the same or different, represent halogen,    (C₁-C₆)hydrocarbyl, optionally substituted with one or two hydroxyl    group or one or more fluorine atoms,-   or, together with the carbon atom to which they are both attached,    R1 and R2 form a (C₃-C₆)carbocyclic ring,-   or one of R1 and R2 taken together with R3 forms a direct bond, such    that a triple bond is constituted,-   or R1 and R2 both represent hydrogen;-   R3 when not forming a direct bond with one of R1 and R2 represents    hydrogen or (C₁-C₃)hydrocarbyl;-   X represents (E)-ethylene, (Z)-ethylene, ethynylene, or a bond;-   Y and Z independently represent hydrogen or methyl; the bond between    C#16 and C#17 is depicted with a dotted line to illustrate that said    bond may be either a single bond, in which case the projection of    the ring substituent is beta, or a double bond;-   A represents hydroxyl, fluorine or hydrogen;-   B represents CH₂ or H₂;-   the configuration in the 3-position corresponds to the same    configuration as in natural vitamin D₃ (normal), or the    configuration in the 3-position is opposite to natural vitamin D₃    (epi);-   with the proviso that when X represents (E)-ethylene or    (Z)-ethylene, one of R1 and R2 taken together with R3 may not form a    direct bond, such that a triple bond is constituted;-   with the further proviso that when X represents a bond R1 and R2 are    not hydrogen;-   with the further proviso that the compound of formula I is not    3(S)-hydroxy-9,10-secocholesta-5(Z),7(E),10(19),22(E),24-penta-ene;-   and prodrugs and stereo isomeric forms thereof.

In the compounds according to formula I, the blocking of the 25-positionis achieved by the presence of a carbon-carbon double- or triple bondbetween carbon #24 and #25. In this way, the 25-position cannot behydroxylated. As discussed more thoroughly later, recent data suggestthat hydroxylation in the 25-position has limited consequences for theparathyroid hormone suppressing effect. Vitamin D analogues which areblocked for hydroxylation in the 25-position therefore retain theirparathyroid hormone suppressing effect while being deprived othervitamin D activities, e.g. the calcemic effect, associated with anintact vitamin D structure.

In another aspect, the invention relates to the use of a compoundaccording to formula I in therapy.

In another aspect, the invention relates to a pharmaceutical compositioncomprising a compound according to formula I.

In still another aspect, the invention relates to methods of treatmentcomprising the step of administering compounds according to formula I toa patient in need thereof.

In a still further aspect, the invention relates to the use of acompound according to formula I in the manufacture of a medicament.

DETAILED DESCRIPTION OF THE INVENTION

In a preferred embodiment of the invention, R1 and R2 when takenseparately, independently represent bromo, chloro, methyl, ethyl,trifluoromethyl, hydroxymethyl, (1- or 2-)hydroxyethyl, normal, iso- orcyclopropyl, 2-hydroxy-9-propyl, 2-methyl-2-propyl, 3-pentyl or3-hydroxy-3-pentyl.

In another preferred embodiment, R1 and R2 are the same, and bothrepresent hydrogen, methyl, ethyl, bromo, chloro, or trifluoromethyl.

In another embodiment R1 and R2 when taken together with the carbon atomto which they are both attached to form a C₃ carbocyclic ring, a C₄carbocyclic ring, a C₅ carbocyclic ring, or a C₆ carbocyclic ring.

In another preferred embodiment, R1 and R2 when taken together includeethylene, tri-methylene, tetra-methylene, or penta-methylene, such as R1and R2 when taken together with the carbon atom to which they are bothattached to form a C₃ carbocyclic ring, a C₄ carbocyclic ring, a C₅carbocyclic ring, or a C₆ carbocyclic ring.

In another preferred embodiment, when R2 constitutes part of a triplebond, then R1 represents a branched C₁₋₆ hydrocarbyl, optionallysubstituted with one or two hydroxyl groups. In particular, R1represents a branched C₁₋₆ hydrocarbyl, optionally substituted with onehydroxyl group, such as —CMe₃, —C(OH)Me₂ or —C(OH)Et₂.

In another preferred embodiment, R3, when not part of a triple bond,represents hydrogen, methyl or cyclopropyl.

In another preferred embodiment A is hydroxyl or fluoro.

In another preferred embodiment R1 and R2 are a radical obtained byremoval of one hydrogen atom from a straight, branched, or cyclicsaturated C₁₋₆ hydrocarbon.

In particular, compounds of formula I may be selected from amongst thelist consisting of

-   1(S),3(R)-Dihydroxy-9,10-secocholesta-5(Z),7(E),10(19),22(E),24-penta-ene    (Compound 1),-   1(S),3(R)-Dihydroxy-9,10-secocholesta-5(Z),7(E),10(19),22(Z),24-penta-ene    (Compound 2),-   20(S),1(S),3(R)-Dihydroxy-9,10-secocholesta-5(Z),7(E),10(19),22(E),24-penta-ene    (Compound 3),-   1(S),3(R)-Dihydroxy-9,10-seco-26,27-cyclo-cholesta-5(Z),7(E),10(19),22(E),24-penta-ene    (Compound 4),-   20(S),1(S),3(R)-Dihydroxy-9,10-seco-26,27-cyclo-cholesta-5(Z),7(E),10(19),22(E),24-penta-ene    (Compound 5),-   1(S),3(R)-Dihydroxy-9,10-seco-26,27-methano-cholesta-5(Z),7(E),10(19),22(E),24-penta-ene    (Compound 6),-   20(S),1(S),3(R)-Dihydroxy-9,10-seco-26,27-methano-cholesta-5(Z),7(E),10(19),22(E),24-penta-ene    (Compound 7),-   1(S),3(R)-Dihydroxy-20(S)-(4,4-dibromo-1,3-butadien-1yl)-9,10-seco-pregna-5(Z),7(E),10(19)-triene    (Compound 8),-   1(S),3(R),26-Trihydroxy-9,10-secocholesta-5(Z),7(E),10(19),22(E),24(E)-penta-ene    (Compound 9),-   20(S),1(S),3(R),26-Trihydroxy-9,10-secocholesta-5(Z),7(E),10(19),22(E),24(E)-penta-ene    (Compound 10),-   1(S),3(R),26-Trihydroxy-9,10-secocholesta-5(Z),7(E),10(19),22(E),24(Z)-penta-ene    (Compound 11),-   20(S),1(S),3(R),26-Trihydroxy-9,10-secocholesta-5(Z),7(E),10(19),22(E),24(Z)-penta-ene    (Compound 12),-   1(S),3(R)-Dihydroxy-20(R)-(4-methyl-5-ethyl-5-hydroxy-1(E),3(E)-heptadienyl)-9,10-secopregna-5(Z),7(E),10(19)-triene    (Compound 13),-   1(S),3(R)-Dihydroxy-20(R)-(3-cyclopropyl-1(E),3-butadienyl)-9,10-secopregna-5(Z),7(E),10(19)-triene    (Compound 14),-   1(S),3(R)-Dihydroxy-9,10-secocholesta-5(Z),7(E),10(19),24-tetra-ene-22-yne    (Compound 15),-   1(S),3(R)-Dihydroxy-20(R)-(5-methyl-5-hydroxy-1,3-hexadiynyl)-9,10-secopregna-5(Z),7(E),10(19)-triene    (Compound 16),-   1(S),3(R)-Dihydroxy-20(S)-(5-ethyl-5-hydroxy-1,3-heptadiynyl)-9,10-secopregna-5(Z),7(E),10(19)-triene    (Compound 17),-   1(S),3(R)-Dihydroxy-20(R)-(5-ethyl-5-hydroxy-1,3-heptadiynyl)-9,10-secopregna-5(Z),7(E),10(19)-triene    (Compound 18),-   1(S),3(R)-Dihydroxy-20(R)-(5,5-dimethyl-1,3-hexadiynyl)-9,10-secopregna-5(Z),7(E),10(19)-triene    (Compound 19),-   1(S),3(R)-Dihydroxy-20(S)-(5,5-dimethyl-1,3-hexadiynyl)-9,10-secopregna-5(Z),7(E),10(19)-triene    (Compound 20),-   1(S)-Fluoro-3(R)-hydroxy-9,10-secocholesta-5(Z),7(E),10(19),22(E),24-penta-ene    (Compound 21),-   1(S),3(R)-Dihydroxy-19-nor-9,10-secocholesta-5,7(E),22(E),24-tetra-ene    (Compound 22),-   1(S),3(S)-Dihydroxy-9,10-secocholesta-5(Z),7(E),10(19),22(E),24-penta-ene    (Compound 23),-   1(S),3(R)-Dihydroxy-9,10-secocholesta-5(Z),7(E),10(19),16,22(E),24-hexa-ene    (Compound 24),-   1(S),3(R)-Dihydroxy-26,26,26,27,27,27-hexafluoro-9,10-secocholesta-5(Z),7(E),10(19),22(E),24-penta-ene    (Compound 25),-   3(S),26-Dihydroxy-9,10-secocholesta-5(Z),7(E),10(19),22(E),24(E)-penta-ene    (Compound 26),-   1(S),3(R)-Dihydroxy-20(R)-(4,4-dibromo-1,3-butadien-1-yl)-9,10-seco-pregna-5(Z),7(E),10(19)-triene    (Compound 27),-   1(S),3(R)-Dihydroxy-26,27-dimethyl-9,10-secocholesta-5(Z),7(E),10(19),22(E),24-penta-ene    (Compound 28),-   1(S),3(S)-Dihydroxy-26,27-dimethyl-9,10-secocholesta-5(Z),7(E),10(19),22(E),24-penta-ene    (Compound 29),-   1(S),3(R)-Dihydroxy-24-methyl-26,27-methano-9,10-secocholesta-5(Z),7(E),10(19),22(E),24-penta-ene    (Compound 30),-   1(S),3(R)-Dihydroxy-20(R)-(4,4-dichloro-1,3-butadien-1-yl)-9,10-seco-pregna-5(Z),7(E),10(19)-triene    (Compound 31),-   1(S),3(R)-Dihydroxy-26,27-ethano-9,10-secocholesta-5(Z),7(E),10(19),22(E),24-penta-ene    (Compound 32),-   1(S),3(R)-Dihydroxy-26,27-propano-9,10-secocholesta-5(Z),7(E),10(19),22(E),24-penta-ene    (Compound 33),-   1(S),3(R)-Dihydroxy-20(S)-cyclopropylidenemethyl-9,10-seco-pregna-5(Z),7(E),10(19)-triene    (Compound 34),-   1(S),3(R)-Dihydroxy-20(R)-cyclopropylidenemethyl-9,10-seco-pregna-5(Z),7(E),10(19)-triene    (Compound 35),-   20(S),1(S),3(R)-Dihydroxy-26,26,26,27,27,27-hexafluoro-9,10-secocholesta-5(Z),7(E),10(19),22(E),24-penta-ene    (Compound 36).

Compounds of formula I may comprise chiral centres, such asasymmetrically substituted carbon atoms, and carbon-carbon double bondswhich give rise to the existence of stereo isomeric forms, such asenantiomers, diastereomers, and geometric isomers (cis/trans). Thepresent invention relates to all such forms, either in pure form or asmixtures thereof.

For example the configuration at C-3 or at C-20 (when Y is differentfrom Z) of formula I can be R or S, or when X is ethylene theconfiguration can be E or Z.

The compounds of formula I may be obtained in crystalline form eitherdirectly by concentration from an organic solvent or by crystallisationor recrystallisation from an organic solvent or mixture of said solventand a cosolvent that may be organic or inorganic, such as water. Thecrystals may be isolated in essentially solvent-free form or as asolvate, such as a hydrate. The invention covers all crystallinemodifications and forms and also mixtures thereof.

In the present context, unless stated differently, the term “prodrug” isintended to indicate compounds in which one or more hydroxyl groups aremasked as groups which can be reconverted to hydroxyl groups in vivo soas to provide compounds of formula I upon administration to a patient.Examples of said groups are esters, e.g carboxylic acid esters andphosphate acid esters. It is well-known that proforms of vitamin D arehydroxylated in the liver and kidneys to reach the biologically activestate. In line with this, compounds of formula I in which A is hydroxylare preferred ones, but compounds in which A is hydrogen are, in fact,another type of prodrug, which may be hydroxylated into an active stateupon administration to a patient.

In the present context, the term “hydrocarbyl” is intended to indicatethe radical obtained by removal of one hydrogen atom from a straight,branched, and/or cyclic, saturated or unsaturated hydrocarbon. Saidstraight, branched, and/or cyclic, saturated or unsaturated hydrocarboninclude but are not limited to methyl, ethyl, 1-propyl, 2-propyl,1-butyl, 2-butyl, 1-pentyl, 2-pentyl, 3-pentyl, 1-hexyl, 2-hexyl,3-hexyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl,2-methyl-2-propyl, 2-methylcyclopropyl, 2-methylallyl, 1-prop-2-ynyl,1-but-2-ynyl, 3-methyl-1-pentyl, 1-hex-1-en-3-ynyl.

In the present context the term “halogen” is intended to indicatecompounds from the seventh main group in the periodic table, i.e.fluoro, chloro, bromo and iodo, and in particular chloro and bromo.

The terms “normal” and “epi” when used to describe the absoluteconfiguration of a compound of the present invention relates to theabsolute configuration of the natural vitamin D₃ itself. Hence, if theconfiguration at a given carbon is referred to as “normal”, itcorresponds to the configuration of vitamin D₃ on that particular carbonatom. Likewise, if the configuration at a given carbon is referred to as“epi”, it is the opposite configuration to that of vitamin D on thatparticular carbon atom.

Of particular relevance for the present invention is the treatment ofsecondary hyperparathyrodism (s-HPT), e.g. in connection with renalfailure, using vitamin D and its analogues. Hyperparathyrodism is adisease characterised by increased secretion of the parathyroid hormonefrom the parathyroid glands. In s-HPT, the cause for the elevatedexcretion is not malfunctioning of the glands, but rather factorsoutside the glands, e.g. failing kidneys. Vitamin D is absorbed from thefood or produced in the skin in a proform which has to be activated toreach its biologically active state. Part of this activation takes placein the kidneys as a hydroxylation of the proform. In patients withfailing kidneys, e.g. dialysis patients, this hydroxylation is impaired,resulting in a lower level of active vitamin D in the blood. Asmentioned above, a low level of vitamin D leads to a high production ofthe parathyroid hormone, and this pathological condition is termedsecondary hyperparathyroidism.

The parathyroid hormone has a powerful influence on the cells in thebones causing them to release calcium into the blood stream. Undernon-pathological conditions this process is well-balanced to secure anadequate calcium level in the bones. However, at elevated parathyroidhormone levels for extended periods of time, the bones will lose toomuch calcium and will become brittle and thus more prone to fracture.This condition is referred to as osteodystrophy and osteomalacia fromwhich renal patients are often suffering. Prolonged exposure toparathyroid hormones is also found to have toxic effects on many vitalorgans, e.g. the heart, skeletal muscles, the nerves and thereproductive system [Malluche, Kidney Int., 367-374, 62, 2002].

One way of controlling the level of parathyroid hormone is to administervitamin D or analogues thereof which will inhibit the secretion of saidhormone. Such therapeutic intervention, however, may be hampered byserious adverse side effects often associated with vitamin D therapy. Asmentioned previously, an effect of vitamin D and many analogues thereofis an increased uptake of calcium from the intestine which may lead tohypercalcemia. This effect may restrict the utility of vitamin Danalogues, which in other respects are beneficial. The aim for much ofthe on-going vitamin D research is thus to minimize the calcemic effectwhile maximizing the clinical effect. Ideally, if the structuralmoieties in the vitamin D molecule responsible for the differentactivities of vitamin D were identified, it would be possible tomanipulate these structures to obtain selectivity, e.g. no calcemicactivity but high parathyroid hormone secretion suppressive effect.Unfortunately, no such clear structure-activity relation has beenestablished yet. However, a recent observation by Brandi in Nephrol DialTransplant, 829-842, 17, 2002 might be helpful in this respect. Brandicompares the PTH suppressive effect of calcitriol, i.e. 1,25(OH)₂ D₃ andits proform, 1α(OH)D₃. 1α(OH)D₃ is hydroxylated in the liver to1,25(OH)₂D₃, and due to the different pharmacokinetics of the twocompounds, the bioavailability of 1,25(OH)₂D₃ was markedly lower when1α(OH)D₃ was administered to the patient than when 1,25(OH)₂ D₃ wasadministered. In spite of this difference in the availability of1,25(OH)₂ D₃ in the two dosing regimes, there was no significantdifference in the suppression of the secretion of PTH. This leads to thespeculation that the 25-hydroxyl group is not mandatory for the PTHsuppressive effect. One way of achieving the desired selectivity couldthus be to block the 25-position in the vitamin D structure so that itcannot be hydroxylated, and in this way maintaining or even increasingthe PTH suppressive effect while dispossessing the molecule of othervitamin D related activities, and in particular its calcemic effect.

The calcemic activities of the compounds of the present invention weredetermined in rats in vivo, as previously described (Binderup, L.,Bramm, E., Biochem. Pharmacol. 37, 889-895 (1988)). The PTH suppressiveeffect of the compounds of the present invention was tested in vitro onbovine parathyroidea cells: Fresh bovine parathyroid glands werecollected from adult cattle within 20 min of slaughter. From mincedparathyroid tissue dispersed parathyroid cells were prepared aspreviously described (E. M. Brown, S. Hurwitz and G. D Aurbach;Preparation of viable isolated bovine parathyroid cells; Endocrinology,1976, vol 99, no 6, 1582-1588). Then, the cells were treated withvitamin D analogues for 60 hours at 37° C. and the PTH secretion wasdetermined.

Table A lists these PTH suppressive effects and the calcemic activitiesof compounds of the present invention. Calcitriol (1,25(OH)₂D₃) is usedas a reference compound.

TABLE A In vivo calcemic activity in rats according to Binderup et al.Biochem. Pharmacol. 37, 889-895 (1988) and the PTH suppressive effecttested in vitro on bovine parathyroidea cells according to the protocolabove of compounds of the present invention relative to Calcitriol(1,25(OH)₂D₃). In vitro PTH secretion relative to untreated In vivocalcemic activity bovine parathyroidea relative to Calcitriol cells(untreated cells Compound (1,25(OH)₂D₃) as control = 100%) Calcitriol100%  60% 1  <10%    57% 3 1% 61% 6 1% 56% 7 <0.5%    65% 25 8% 74% 27<0.5%    65% 28 1% 62%

The data listed in Table A show that the compounds of the presentinvention have a comparable PTH suppressive effect as the referencecompound Calcitriol (1,25(OH)₂D₃), together with a surprisingly lowercalcemic effect than the reference compound.

The compounds of the present invention thus surprisingly show a highselectivity for e.g. the inhibition of the production of theparathytoidea hormone without the unwanted effects of vitamin Danalogues, such as 1,25(OH)₂D₃, on the calcium balance in the organism,including calcemic and calciuric activities.

In a particular embodiment, the invention thus provides a method fortreating, preventing or ameliorating s-HPT, and in particular s-HPTassociated with renal failure, the method comprising administering to apatient in need thereof an effective amount of a compound of formula I.Optionally, said method may include treatment with other therapeuticallyactive compounds normally used in the treatment of the above mentioneddisease. Said compounds may be administered concomitantly orsequentially with compounds of the present invention, and in theyparticular include phosphate binders.

The use of compounds of the present invention may not be limited to thetreatment of s-HPT. It is well-known that vitamin D and analoguesthereof may be beneficial in the treatment of a variety of diseases dueto a strong activity in inducing differentiation and inhibitingundesirable proliferation of certain cells, including skin cells andcancer cells, as well as an immunomodulating effect and an effect inbone build-up and maintenance (Brown A J: Vitamin D analogues. Am JKidney Dis 32 (Suppl): S25-S39, 1998; Brown A J et al.: Vitamin D. AmPhysiol 277:F157-F175, 1999). Accordingly, the invention also provides amethod of treating, preventing or ameliorating diseases characterised byabnormal cell differentiation and/or cell proliferation, cancer,leukemia, mammary cancer, brain glial cancer, osteosarcoma, melanoma,myelofibrosis, psoriasis, primary hyperparathyroidism, diabetes melitus,discoid and systemic lupus erythematosus, chronic dermatoses ofautoimmune type, hypertension, acne, alopecia, skin aging, AIDS,neurodegenerative disorders, Alzheimer's disease, host versus graft andgraft versus host reactions, rejections of transplants, steroid inducedskin atrophy and osteroporosis, and for inducing osteogenesis, themethod comprising administering to a patient in need thereof aneffective amount of a compound of formula I. Optionally, said method mayinclude treatment with other therapeutically active compounds normallyused in the treatment of the above mentioned diseases. Said compoundsmay be administered concomitantly or sequentially with compounds of thepresent invention, and they include phosphate binders, steroids andanti-proliferative agents.

In the systemic treatment using the present invention daily doses offrom 0.001-2 μg per kilogram body weight, preferably from 0.002-0.3μg/kg of mammal body weight, for example 0.003-0.3 μg/kg of a compoundof formula I is administered, typically corresponding to a daily dosefor an adult human of from 0.1 to 200 μg. A suitable dosing regime may,however, also include dosing with longer intervals, e.g. every otherday, every week, or even with longer intervals. In the topical treatmentof dermatological disorders, ointments, creams or lotions containingfrom 0.1-1000 μg/g, and preferably from 0.1-500 μg/g, for example0.1-200 μg/g of a compound of formula I is administered. For topical usein ophthalmology ointments, drops or gels containing from 0.1-1000 μg/g,and preferably from 0.1-500 μg/g, for example 0.1-100 μg/g of a compoundof formula I is administered. The oral compositions are formulated,preferably as tablets, capsules, or drops, containing from 0.07-100 μg,preferably from 0.1-50 μg, of a compound of formula I per dosage unit.

In a further preferred aspect, the invention relates to a pharmaceuticalcomposition comprising a compound of formula I. The formulations of thepresent invention, both for veterinary and for human medical use,comprise active ingredients in association with a pharmaceuticallyacceptable carrier(s) and optionally other therapeutic ingredient(s).The carrier(s) must be “acceptable” in the sense of being compatiblewith the other ingredients of the formulations and not deleterious tothe recipient thereof.

Conveniently, dosage unit of a formulation contain between 0.05 μg and100 μg, preferably between 0.1 μg and 50 μg of a compound of formula I.

By the term “dosage unit” is meant a unitary, i.e. a single dose whichis capable of being administered to a patient, and which may be readilyhandled and packed, remaining as a physically and chemically stable unitdose comprising either the active material as such or a mixture of itwith solid or liquid pharmaceutical diluents or carriers.

The formulations include e.g. those in a form suitable for oral(including sustained or timed release), rectal, parenteral (includingsubcutaneous, intraperitoneal, intramuscular, intraarticular andintravenous), transdermal, ophthalmic, topical, nasal or buccaladministration.

The formulations may conveniently be presented in dosage unit form andmay be pre-pared by any of the methods well known in the art ofpharmacy, e.g as disclosed in Remington, The Science and Practice ofPharmacy, 20^(th) ed., 2000. All methods include the step of bringingthe active ingredient into association with the carrier, whichconstitutes one or more accessory ingredients. In general, theformulations are prepared by uniformly and intimately bringing theactive ingredient into association with a liquid carrier or a finelydivided solid carrier or both, and then, if necessary, shaping theproduct into the desired formulation.

Formulations of the present invention suitable for oral administrationmay be in the form of discrete units as capsules, sachets, tablets orlozenges, each containing a predetermined amount of the activeingredient; in the form of a powder or granules; in the form of asolution or a suspension in an aqueous liquid or non-aqueous liquid,such as ethanol or glycerol; or in the form of an oil-in-water emulsionor a water-in-oil emulsion. Such oils may be edible oils, such as e.g.cottonseed oil, sesame oil, coconut oil or peanut oil. Suitabledispersing or suspending agents for aqueous suspensions includesynthetic or natural gums such as tragacanth, alginate, acacia, dextran,sodium carboxymethylcellulose, gelatin, methylcellulose,hydroxypropylmethylcellulose, hydroxypropylcellulose, carbomers andpolyvinylpyrrolidone. The active ingredients may also be administered inthe form of a bolus, electuary or paste.

A tablet may be made by compressing or moulding the active ingredientoptionally with one or more accessory ingredients. Compressed tabletsmay be prepared by compressing, in a suitable machine, the activeingredient(s) in a free-flowing form such as a powder or granules,optionally mixed by a binder, such as e.g. lactose, glucose, starch,gelatine, acacia gum, tragacanth gum, sodium alginate,carboxymethylcellulose, methylcellulose, hydroxypropylmethylcellulose,polyethylene glycol, waxes or the like; a lubricant such as e.g. sodiumoleate, sodium stearate, magnesium stearate, sodium benzoate, sodiumacetate, sodium chloride or the like; a disintegrating agent such ase.g. starch, methylcellulose, agar, bentonite, croscarmellose sodium,sodium starch glycollate, crospovidone or the like Or a dispersingagent, such as polysorbate 80. Moulded tablets may be made by moulding,in a suitable machine, a mixture of the powdered active ingredient andsuitable carrier moistened with an inert liquid diluent.

Formulations for rectal administration may be in the form ofsuppositories in which the compound of the present invention is admixedwith low melting water soluble or insoluble solids such as cocoa butter,hydrogenated vegetable oils, polyethylene glycol or fatty acids estersof polyethylene glycols, while elixirs may be prepared using myristylpalmitate.

Formulations suitable for parenteral administration convenientlycomprise a sterile oily or aqueous preparation of the activeingredients, which is preferably isotonic with the blood of therecipient, e.g. isotonic saline, isotonic glucose solution or buffersolution. The formulation may be conveniently sterilised by for instancefiltration through a bacteria retaining filter, addition of sterilisingagent to the formulation, irradiation of the formulation or heating ofthe formulation. Liposomal formulations as disclosed in e.g.Encyclopedia of Pharmaceutical Technology, vol. 9, 1994, are alsosuitable for parenteral administration.

Alternatively, the compound of formula I may be presented as a sterile,solid preparation, e.g. a freeze-dried powder, which is readilydissolved in a sterile solvent immediately prior to use.

Transdermal formulations may be in the form of a plaster or a patch.

Formulations suitable ophthalmic administration may be in the form of asterile aqueous preparation of the active ingredients, which may be inmicrocrystalline form, for example, in the form of an aqueousmicrocrystalline suspension. Liposomal formulations or biodegradablepolymer systems e.g. as disclosed in Encyclopedia of PharmaceuticalTechnology, vol. 2, 1989, may also be used to present the activeingredient for ophthalmic administration.

Formulations suitable for topical or ophthalmic administration includeliquid or semi-liquid preparations such as liniments, lotions, gels,applicants, oil-in-water or water-in-oil emulsions such as creams,ointments or pastes; or solutions or suspensions such as drops.

Formulations suitable for nasal or buccal administration include powder,self-propelling and spray formulations, such as aerosols and atomisers.

Prodrugs of the present invention may also be delivered by use ofmonoclonale antibodies as individual carriers to which the compoundmolecules are coupled.

In addition to the aforementioned ingredients, the formulations of acompound of formula I may include one or more additional ingredientssuch as diluents, buffers, flavouring agents, colourant, surface activeagents, thickeners, preservatives, e.g. methyl hydroxybenzoate(including anti-oxidants), emulsifying agents and the like.

Furthermore, said formulations may also comprise other therapeuticallyactive compounds normally used in the treatment of the above mentioneddiseases. Examples of such compounds include phosphate binders, steroidsand anti-proliferative agents.

In still another aspect, the invention relates to the use of a compoundof formula I, optionally together with another therapeutically activecompound in the manufacture of a medicament intended for the treatmentof abnormal cell differentiation and/or cell proliferation, cancer,leukemia, mammary cancer, brain glial cancer, osteosarcoma, melanoma,myelofibrosis, psoriasis, primary hyperparathyroidism, s-HPT, s-HPT inassociation with renal failure, diabetes melitus, discoid and systemiclupus erythematosus, chronic dermatoses of autoimmune type,hypertension, acne, alopecia, skin aging, AIDS, neurodegenerativedisorders, Alzheimer's disease, host versus graft and graft versus hostreactions, rejections of transplants, steroid induced skin atrophy andosteroporosis, and for inducing osteogenesis. Said other therapeuticallyactive compound may conveniently be selected from amongst, e.g.phosphate binders, steroids and anti-proliferative agents.

A compound of formula I may be prepared from the intermediates 1according to the reaction Scheme 1.

The symbol * is used in this Scheme to indicate that the group A* in anintermediate compound may either be identical to the group A as found incompound I (for example, fluorine in N_(Z)), or alternatively may be agroup that can be converted to this at any subsequent stage in thesynthesis (for example, silyl ether protected hydroxyl in N_(Z)).Although not formally indicated in this manner, the same situation mayalso apply for the variables R¹, R², R³, Y, and Z on Scheme 1.Furthermore, the identity of N (i.e. N_(E), N_(Z), or N_(CD)) and/or ofone or more variable group(s) may change from intermediate tointermediate along the reaction sequence. However the actual identitywill be apparent from the particular context. Note that while Cl and Liare atomic symbols (as are C, H, and O), the letter W is used as anabbreviation for a functional group or element that stabilises theorgano-lithium species. The configuration (E or Z) of the double bond(to become X=ethylene in the final compounds I) is left unspecified inScheme 1 (by drawing a linear arrangement) but is specified as and whenrequired.

Starting materials of type 1 and certain examples of intermediates oftype 2, 4, 6, 7, and 8 are well known to a person skilled in the art andhave been described in the literature such as by G.-D. Zhu and W. H.Okamura in Chem. Rev. 1995, 95, 1877-1952. The suffixes, a, b and c,identify the structure of the N group, as defined at the top of Scheme1.

In outline, the suggested reactions, all well known to the syntheticorganic chemist skilled in the art of vitamin D chemistry, are carriedout as follows. Standard abbreviations are used throughout thisdisclosure, e.g. Ac=acetyl; aq.=aqueous; DCM=dichloromethane; Et=ethyl;ether=diethylether; h=hour(s); LDA=lithium diisopropylamide; Me=methyl;PDC=pyridinium dichromate; TBA=tetra(n)butylammonium;TBS=t-butyldimethylsilyl; TMS=trimethylsilyl; Ts=p-tosyl;DIBAL=diisobutylaluminium hydride; Ph=phenyl; THF=tetrahydrofuran;v=volume.

1→2 Wittig or Wadsworth-Emmons reaction [e.g. Wittig with Ph₃P═CH—C(O)R³(for R3=H, via the R3=OMe intermediate, from which it is derived bysequential DIBAL reduction and PDC or Dess-Martin periodinaneoxidation).] The configuration of the double bond established in thisreaction is usually E (small amounts of the Z intermediate can often beisolated however), but conditions can be selected to give an increasedproportion of the Z intermediate, e.g. a modification of theWadsworth-Emmons reaction using (CF₃CH₂O)₂P(O)—CH₂C(O)OMe. TheE-configuration can alternatively be converted to Z byphotoisomerisation at the stage of intermediate 2. Separation of therequired isomer from a mixture of E and Z isomers can be performed atthis or a later convenient stage by e.g. chromatography orcrystallisation.

-   1→4 For intermediates with W═S(O₂)Ph or W=SeMe, methods have been    described in the literature, such as by M. J. Calverly, Tedrahedron    Letters 1987, 28, 1337-1340.-   1→5 E.g. by Wittig reaction with Ph₃P═CH—C(R³)═CR¹R². E and Z    isomers formed in this reaction can optionally be separated by e.g.    chromatography or crystallisation at this stage or any later stage    in the synthesis.-   1→6→7→8→7 The dibromo-intermediate may be used instead of the shown    dichloro-intermediate (6) in this part of a well-known reaction    sequence for making alkynes as described earlier by e.g. W. G.    Salmond et al., Tetrahedron Letters 1977, 14, 1239-1240.-   2→3 For R³═H (in 3): Sequential DIBAL reduction [for R³═OMe (or H)    in 2 (see 1→2), conversion of the alcohol to a leaving group (e.g.    Cl or OTs) which is then substituted to incorporate W (e.g. —P(O)Ph2    or —S(O₂)Ph, either directly or via oxidation of the lower oxidation    state —PPh2 or —SPh, all available as salts), and lithiation (e.g.    with n-BuLi or LDA).-   2→5 Wittig reaction e.g. with Ph₃P═CR¹R² or Wadsworth-Emmons    reaction e.g. using (EtO)₂P(O)—CR¹R². The separation of 24-E and    24-Z isomers formed in this reaction can can optionally be separated    by e.g. chromatography or crystallisation at this stage or any later    stage in the synthesis.-   3→5 Coupling reaction with a carbonyl compound, e.g. by a Horner    reaction when W═P(O)Ph2, or by a Julia reaction when W═S(O₂)Ph    (followed in the latter reaction by a reductive elimination of W    together with the oxy-group). The separation of 24-E and 24-Z    isomers formed can optionally be separated by e.g. chromatography or    crystallisation at this stage or any later stage in the synthesis.-   4→5 Coupling reaction with a carbonyl compound followed by    elimination of W and the oxy-group. Separation of the required    isomer from a mixture of E and Z isomers can be performed at this or    a later convenient stage.-   7→9 Coupling reaction with a carbonyl compound.-   8→10 Palladium catalysed cross coupling with a terminal acetylene or    with a vinyl or acetylenic derivative such as a halogenide (e.g.    bromide). The reactions include, but are not limited, to Heck,    Suzuki, Cadiot-Chodkiewski; Negishi, Sonogashira, and Stille type    reactions.-   9→10 Dehydration with Martin's sulfurane reagent.-   5 or 10→I “N→M”: see below. In addition A*, R¹, R², R³, Y, and/or Z    may be transformed or derivatised by methods and general procedures    well known to a person skilled in the art, such as described in    “Comprehensive Organic Transformations”, by R. C. Larock, VCH 1989.-   N_(CD)→M: Sequential desilylation with HF to give the alcohol,    oxidation with Dess-Martin periodinane to the ketone, and    Horner-Wittig coupling with the lithio-derivative of the requisite    known A-ring phosphine oxide of formula II to give N_(Z). Then    desilylation with HF or TBA-fluoride.

-   N_(E)→M: Conversion to N_(Z) (B═CH₂) by triplet-sensitised    photoisomerisation, then desilylation with HF or TBA-fluoride.-   N_(Z)→M: Desilylation with HF or TBA-fluoride.

The invention is further illustrated by the following Preparations andExamples: The exemplified compounds according to formula I are listed inTable 1, whereas the starting materials and intermediates of generalformulae 1 through 10 (Scheme 1) are listed in Table 2.

TABLE 1 Example General Compound Number Procedure A B 3-Configuration16,17-bond X Y Z R1 R2 R3 1 1 9 OH CH₂ normal single E-ethylene H Me MeMe H 2 2 9 OH CH₂ normal single Z-ethylene H Me Me Me H 3 3 9 OH CH₂normal single E-ethylene Me H Me Me H 4 4 8 OH CH₂ normal singleE-ethylene H Me —(CH₂)₂— H 5 5 9 OH CH₂ normal single E-ethylene Me H—(CH₂)₂— H 6 6 9 OH CH₂ normal single E-ethylene H Me —(CH₂)₃— H 7 7 9OH CH₂ normal single E-ethylene Me H —(CH₂)₃— H 8 18 8 OH CH₂ normalsingle E-ethylene Me H Br Br H 9 8 9 OH CH₂ normal single E-ethylene HMe Me CH₂OH H 10 9 9 OH CH₂ normal single E-ethylene Me H Me CH₂OH H 1110 9 OH CH₂ normal single E-ethylene H Me CH₂OH Me H 12 11 9 OH CH₂normal single E-ethylene Me H CH₂OH Me H 13 12 9 OH CH₂ normal singleE-ethylene H Me Me C(OH)Et₂ H 14 13 9 OH CH₂ normal single E-ethylene HMe H H cyclopropyl 15 14 8 OH CH₂ normal single ethynylene H Me Me Me H16 15 8 OH CH₂ normal single ethynylene Me H C(OH)Me₂ bond bond 17 16 8OH CH₂ normal single ethynylene H Me C(OH)Et₂ bond bond 18 17 8 OH CH₂normal single ethynylene Me H C(OH)Et₂ bond bond 19 OH CH₂ normal singleethynylene H Me CMe₃ bond bond 20 OH CH₂ normal single ethynylene Me HCMe₃ bond bond 21 19 F CH₂ normal single E-ethylene H Me Me Me H 22 20OH H₂ normal single E-ethylene H Me Me Me H 23 21 OH CH₂ epi singleE-ethylene H Me Me Me H 24 22 OH CH₂ normal double E-ethylene H Me Me MeH 25 23 8 OH CH₂ normal single E-ethylene H Me CF₃ CF₃ H 26 H CH₂ normalsingle E-ethylene H Me Me CH₂OH H 27 25 8 OH CH₂ normal singleE-ethylene H Me Br Br H 28 26 8 OH CH₂ normal single E-ethylene H Me EtEt H 29 27 8 OH CH₂ epi single E-ethylene H Me Et Et H 30 28 9 OH CH₂normal single E-ethylene H Me —(CH₂)₃— Me 31 29 8 OH CH₂ normal singleE-ethylene H Me Cl Cl H 32 30 9 OH CH₂ normal single E-ethylene H Me—(CH₂)₄— H 33 31 9 OH CH₂ normal single E-ethylene H Me —(CH₂)₅— H 34 329 OH CH₂ normal single bond Me H —(CH₂)₂— H 35 33 9 OH CH₂ normal singlebond H Me —(CH₂)₂— H 36 36 8 OH CH₂ normal single E-ethylene Me H CF₃CF₃ H

TABLE 2 Eth- Com- Gen- 3-Con- ylene pound Type Prepar- eral fig- con-En- num- (Scheme ation Proce- ura- 16,17- figu- try ber 1) Number dureA* B tion bond Y Z ration R1 R2 R3 W 1 101 1a O-TBS normal single H Me 2102 1c single H Me 3 103 1a O-TBS normal single Me H 4 104 1a O-TBSnormal double H Me 5 202 2a O-TBS normal single H Me H 6 203 2a O-TBSnormal single Me H H 7 204 2a O-TBS normal single H Me E cyclo- propyl 8205 2a 35 O-TBS normal single H Me E Me 9 206 2c single H Me H 10 301 3aO-TBS normal single H Me E H SO₂—Ph 11 302 3a 33 O-TBS normal single HMe E H P(O)Ph₂ 12 401 4a O-TBS normal single H Me H SO₂—Ph 13 402 4aO-TBS normal single H Me H P(O)Ph₂ 14 403 4a O-TBS normal single H Me HSeMe 15 501 5a 1 1 O-TBS normal single H Me E Me Me H 16 502 5a 8 O-TBSnormal single H Me Z Me Me H 17 503 5a 3 1 O-TBS normal single Me H E MeMe H 18 504 5a 4 1 O-TBS normal single H Me E —(CH₂)₂— H 19 505 5a 5 1O-TBS normal single Me H E —(CH₂)₂— H 20 506 5a 6 1 O-TBS normal singleH Me E —(CH₂)₃— H 21 507 5a 7 1 O-TBS normal single Me H E —(CH₂)₃— H 22508 5a 9 2 O-TBS normal single H Me E Me CO₂Et H 23 509 5a 9 2 O-TBSnormal single H Me E CO₂Et Me H 24 510 5a 11 3 O-TBS normal single H MeE Me CH₂OH H 25 511 5a 12 3 O-TBS normal single H Me E CH₂OH Me H 26 5125a 10 2 O-TBS normal single Me H E Me CO₂Et H 27 513 5a 10 2 O-TBSnormal single Me H E CO₂Et Me H 28 514 5a 13 3 O-TBS normal single Me HE Me CH₂OH H 29 515 5a 14 3 O-TBS normal single Me H E CH₂OH Me H 30 5165a 15 4 O-TBS normal single H Me E Me C(OH)Et₂ H 31 517 5a 16 O-TBSnormal single H Me E H H cyclo- propyl 32 518 5a 17 O-TBS normal singleMe H E Br Br H 33 534 5a 34 O-TBS normal single H Me E CF₃ CF₃ H 34 5195b 101 7 O-TBS CH₂ normal single H Me E Me Me H 35 520 5b 114 7 O-TBSCH₂ normal single Me H E Br Br H 36 521 5b 102 7 O-TBS CH₂ normal singleH Me Z Me Me H 37 522 5c 2 1 single H Me E Me Me H 38 523 5b 103 7 O-TBSCH₂ normal single Me H E Me Me H 39 524 5b 104 7 O-TBS CH₂ normal singleH Me E —(CH₂)₂— H 40 525 5b 105 7 O-TBS CH₂ normal single Me H E—(CH₂)₂— H 41 526 5b 106 7 O-TBS CH₂ normal single H Me E —(CH₂)₃— H 42527 5b 107 7 O-TBS CH₂ normal single Me H E —(CH₂)₃— H 43 528 5b 108 7O-TBS CH₂ normal single H Me E Me CH₂OH H 44 529 5b 109 7 O-TBS CH₂normal single H Me E CH₂OH Me H 45 530 5b 110 7 O-TBS CH₂ normal singleMe H E Me CH₂OH H 46 531 5b 111 7 O-TBS CH₂ normal single Me H E CH₂OHMe H 47 532 5b 112 7 O-TBS CH₂ normal single H Me E Me C(OH)Et₂ H 48 5335b 113 7 O-TBS CH₂ normal single H Me E H H cyclo- propyl 49 535 5b 1287 O-TBS CH₂ normal single H Me E CF₃ CF₃ H 50 601 6a O-TBS normal singleH Me 51 602 6a O-TBS normal single Me H 52 701 7a 18, 19 5 O-TBS normalsingle H Me 53 702 7a 20 5 O-TBS normal single Me H 54 801 8a 19 5 O-TBSnormal single H Me 55 802 8a 20 5 O-TBS normal single Me H 56 901 9a 185 O-TBS normal single H Me Me Me H 57 1001 10a  18 5 O-TBS normal singleH Me Me Me H 58 1002 10b  115 7 O-TBS CH₂ normal single H Me Me Me H 591003 10a  21 6 O-TBS normal single Me H C(OH)Me₂ bond bond 60 1004 10b 116 7 O-TBS CH₂ normal single Me H C(OH)Me₂ bond bond 61 1005 10a  22 6O-TBS normal single H Me C(OH)Et₂ bond bond 62 1006 10b  117 7 O-TBS CH₂normal single H Me C(OH)Et₂ bond bond 63 1007 10a  23 6 O-TBS normalsingle Me H C(OH)Et₂ bond bond 64 1008 10b  118 7 O-TBS CH₂ normalsingle Me H C(OH)Et₂ bond bond 65 1009 10a  6 O-TBS normal single H MeCMe₃ bond bond 66 1010 10b  7 O-TBS CH₂ normal single H Me CMe₃ bondbond 67 1011 10a  6 O-TBS normal single Me H CMe₃ bond bond 68 1012 10b 7 O-TBS CH₂ normal single Me H CMe₃ bond bond 69 536 5a 24 O-TBS normalsingle H Me E Br Br H 70 538 5a 25 O-TBS normal single H Me E Cl Cl H 71540 5a 26 O-TBS normal single H Me E —(CH₂)₃— Me 72 542 5a 27 1 O-TBSnormal single H Me E —(CH₂)₄— H 73 544 5a 28 1 O-TBS normal single H MeE —(CH₂)₅— H 74 546 29 1 O-TBS normal single Me H bond —(CH₂)₂— H 75 54830 1 O-TBS normal single H Me bond —(CH₂)₂— H 76 550 5a 31 O-TBS normalsingle H Me E Et Et H 77 303 5b 32 O-TBS CH₂ epi single H Me E P(O)Ph₂78 537 5b 119 7 O-TBS CH₂ normal single H Me E Br Br H 79 539 5b 120 7O-TBS CH₂ normal single H Me E Cl Cl H 80 541 5b 121 7 O-TBS CH₂ normalsingle H Me E —(CH₂)₃— Me 81 543 5b 122 7 O-TBS CH₂ normal single H Me E—(CH₂)₄— H 82 545 5b 123 7 O-TBS CH₂ normal single H Me E —(CH₂)₅— H 83547 124 7 O-TBS CH₂ normal single Me H bond —(CH₂)₂— H 84 549 125 7O-TBS CH₂ normal single H Me bond —(CH₂)₂— H 85 551 5b 126 7 O-TBS CH₂normal single H Me E Et Et H 86 552 5b 127 O-TBS CH₂ epi single H Me EEt Et H

PREPARATIONS AND EXAMPLES

Reactions were routinely (unless otherwise noted) run by stirring underan argon atmosphere, with additions of reagent (liquid or in solution)occurring drop wise via a syringe. As standard work-up procedure, theorganic layer was separated, washed sequentially with water andsaturated sodium chloride solution, dried over anhydrous magnesiumsulphate, and concentrated in vacuo to give a crude product, which wasthen purified by chromatography. All preparative and analytical (TLC)chromatography was performed on silica gel using a gradient from 1% to50% (v:v) ether (i.e. diethyl ether) in petroleum ether as eluent, orfrom 30% (v:v) ethyl acetate in petroleum ether to pure ethyl acetate.In the General Procedures, the variable entries are indented (onseparate lines) and then listed in the specific Preparations, togetherwith details if needed on any deviations from the General Procedure thatwere actually employed. However the proportional scaling of thequantities of non-variable reagents and solvents to the molar quantitiesspecified in each Preparation is taken for granted and thus notconsidered a deviation requiring explicit details.

Compounds were characterised spectroscopically: For ¹H nuclear magneticresonance spectra (300 MHz) and ¹³C NMR (75.6 MHz) chemical shift values(δ) (in ppm) are quoted, unless otherwise specified, fordeuteriochloroform solutions relative to internal tetramethylsilane(added, ¹H-NMR: δ=0.00 ppm, ¹³C-NMR: δ=0.00 ppm) or chloroform(residual, ¹H-NMR: δ=7.25 ppm) or deuteriochloroform (¹³C-NMR: δ=76.81ppm). The value for a multiplet (¹H-NMR), either defined [doublet (d),triplet (t), quartet (q)] or not (m) at the approximate mid point isgiven unless a range is quoted (s=singlet, b=broad). In some cases, onlyselected, characteristic signals may be reported for the intermediatecompounds i.e. those of Table 2.

General Procedure 1 (Preparations 1-7 and 27-28) [2→5] (Preparations29-30) [1→5]

To a solution or suspension, maintained at about −70° C., of analkyl-triphenylphosphonium salt (7 mmol) in dry THF (50 ml) was addedn-butyllithium (1.6M in hexanes, 4.36 ml, 7 mmol). The temperature ofthe mixture was then allowed to rise to 0° C. for 20 min, after whichrecooling at −70° C. was resumed for the addition of intermediate 1 or 2(4 mmol), dissolved in dry THF (8 ml). After 30 minutes at the sametemperature, slow warming up, and 70 min at room temperature, themixture was partitioned between saturated ammonium chloride solution andethyl acetate, and worked up according to the standard work-up procedureabove to give intermediate 5.

Preparation 1: Compound 501

Alkyl-triphenylphosphonium salt: Isopropyl-triphenylphosphonium iodide(3.16 g, 7.3 mmol).

Intermediate 2: 202 (2.39 g).

Purification of compound 502 from the crude product by directcrystallisation from methanol, omitting the chromatography step:¹³C-NMR: δ=153.5, 143.1, 138.1, 135.2, 132.5, 125.1, 124.1, 121.5,116.3, 106.4, 70.0, 67.1, 56.3, 56.2, 45.7, 43.8, 40.4, 40.3, 36.4,28.8, 27.7, 25.7, 25.6, 23.3, 22.0, 20.6, 18.1, 18.0, 17.9, 12.1, −4.9,−5.1 ppm.

Preparation 2: Compound 522

Alkyl-triphenylphosphonium salt: Isopropyl-triphenylphosphonium iodide(1.55 g, 3.6 mmol).

Intermediate 2: 206 (Prepared from 102 analogously to the describedpreparation of 202 from 101 in WO9100855) (0.7 g, 2 mmol).

Compound 522: ¹³C-NMR: δ=138.5, 132.3, 125.1, 123.8, 69.3, 56.4, 52.9,42.0, 40.5, 39.7, 34.3, 29.2, 25.7, 25.6, 22.8, 20.3, 18.0, 17.8, 17.5,13.8, −5.0, −5.3 ppm.

Preparation 3: Compound 503

Alkyl-triphenylphosphonium salt: Isopropyl-triphenylphosphonium iodide(1.47 g, 3.4 mmol).

Intermediate 2: 203 (1 g, 1.67 mmol).

Purification of compound 503 by chromatography (2% v:v, ether inpetroleum ether).

Compound 503: ¹³C-NMR: δ=153.4, 143.3, 138.5, 135.1, 132.3, 125.1,124.0, 121.6, 116.1, 106.4, 70.1, 67.0, 56.7, 56.1, 45.8, 43.8, 40.6,39.6, 36.4, 28.8, 27.1, 25.7, 25.7, 25.6, 23.3, 21.9, 21.3, 18.1, 17.9,12.1, −4.9, −5.1 ppm.

Preparation 4: Compound 504

Alkyl-triphenylphosphonium salt: Cyclopropyl-triphenylphosphoniumbromide (0.76 g, 2 mmol).

Intermediate 2: 202 (0.60 g, 1 mmol)

Purification of compound 504 by chromatography (5% v:v, ether inpetroleum ether).

Compound 504: ¹³C-NMR: δ=6.45 (d, J=12 Hz, 1H), 6.33 (bd, J=10 Hz, 1H),6.12 (dd, J=15 Hz, J=10 Hz, 1H), 5.81 (d, J=12 Hz, 1H), 5.55 (dd, J=9Hz, J=15 Hz, 1H), 4.97 (m, 1H), 4.93 (m, 1H), 4.53 (m, 1H), 4.23 (m,1H), 2.86 (m, 1H), 2.55 (dd, 1H), 2.30 (bd, 1H), 2.20-1.15 (m, 14H),1.15-1.0 (m, 14H), 1.07 (d, J=7 Hz, 3H), 0.89 (s, 9H), 0.86 (s, 9H),0.57 (s, 3H), 0.05 (m, 12H) ppm.

Preparation 5: Compound 505

Alkyl-triphenylphosphonium salt: Cyclopropyl-triphenylphosphoniumbromide (1.53 g, 4 mmol).

Intermediate 2: 203 (0.56 g, 0.93 mmol)

Purification of compound 505 by chromatography (5% v:v, ether inpetroleum ether).

Compound 505: ¹³C-NMR: δ=6.44 (d, J=11.4 Hz, 1H), 6.35 (bd, J=10.3 Hz,1H), 6.08 (dd, J=10.3 Hz, J=15.3, 1H), 5.81 (d, J=11.4 Hz, 1H), 5.55(dd, J=9.5 Hz, J=15.3, 1H), 4.97 (bt, 1H), 4.93 (bt, 1H), 4.52 (m, 1H),4.21 (m, 1H), 2.84 (m, 1H), 2.54 (dd, J=5.3 Hz, J=14.5 Hz, 1H), 2.31(bd, J=13.7 Hz, 1H), 2.2-1.0 (m, 18H), 0.97 (d, J=6.5 Hz, 3H), 0.89 (s,9H), 0.86 (s, 9H), 0.52 (s, 3H), 0.05 (s, 12H) ppm.

Preparation 6: Compound 506

Alkyl-triphenylphosphonium salt: Cyclobutyl-triphenylphosphonium bromide(0.80 g, 2 mmol).

Intermediate 2: 202 (0.60 g, 1 mmol)

Purification of compound 506 by chromatography (1% v:v, ether inpetroleum ether).

Compound 506: ¹³C-NMR: δ=153.5, 143.1, 142.5, 137.1, 135.2, 123.6,121.5, 120.8, 116.3, 106.4, 70.0, 67.0, 56.3, 56.2, 45.7, 43.8, 40.3,40.3, 36.4, 31.1, 29.7, 28.7, 27.7, 25.7, 25.6, 23.3, 22.0, 20.5, 18.1,17.9, 17.0, 12.0, −4.9, −5.0, −5.1 ppm.

Preparation 7: Compound 507

Alkyl-triphenylphosphonium salt: Cyclobutyl-triphenylphosphonium bromide(1.33 g, 3.34 mmol).

Intermediate 2: 203 (1 g, 1.67 mmol).

Purification of compound 507 by chromatography (1% v:v, ether inpetroleum ether).

Chromatography: 1% ether in petroleum ether.

Compound 507: ¹³C-NMR: δ=153.4, 143.3, 142.4, 137.4, 135.1, 123.6,121.6, 120.9, 116.1, 106.4, 70.1, 67.0, 56.8, 56.1, 45.8, 43.8, 40.6,39.6, 36.4, 31.1, 29.7, 28.8, 27.1, 25.7, 25.6, 23.3, 21.9, 21.3, 18.1,17.9, 16.9, 12.1, −4.9, −5.1 ppm.

Preparation 8: Compound 502 [4→5]

To a solution, maintained at about −70° C., of the lithio-derivative 403(Table 2., entry 14) (prepared from 0.75 g, 1 mmol of the seleno-acetalderivative of 101 in dry THF (5 ml)) was added dropwise the side chainbuilding block 3-methyl-crotonaldehyde (0.10 g, 1.2 mmol). Afterstirring at the same temperature for 30 min, the reaction was quenchedwith wet THF and the mixture partitioned between ether and water and togive the intermediate adduct as a mixture of diastereoisomers. To asolution of this, maintained at about 5° C., in dry DCM (15 ml) andtriethylamine (2 ml) was added methanesulphonyl chloride (1.5 mmol).After stirring at the same temperature for 30 min, the reaction mixturewas partitioned between ether and water. The standard work-up procedureabove gave compound 502: ¹³C-NMR: δ=153.5, 143.1, 136.0, 135.2, 134.4,121.7, 121.5, 120.4, 116.3, 106.4, 70.0, 67.0, 56.5, 56.3, 45.7, 43.8,40.4, 40.3, 36.4, 28.8, 27.3, 25.7, 25.6, 23.4, 22.0, 20.6, 18.0, 17.9,12.2, −4.9, −5.0, −5.1 ppm.

General Procedure 2 (Preparations 9, 10) [2→5]

To a solution, maintained at about 20° C., of Intermediate 2 (0.76 mmol)in DCM (5 ml) was added triethyl 2-phosphonopropionate (0.45 g, 1.9mmol), 50% aqueous NaOH solution (5 ml), and TBA hydrogen sulphate (0.11g). After vigorous stirring for 105 min, the reaction mixturepartitioned between water and ethyl acetate. Standard work-up as above(chromatography 2%-5% v:v ethyl acetate in petroleum ether as eluent)gave Intermediates 5.

Preparation 9: Compound 508 and 509

Intermediate 2: 202 (0.46 g).

First eluted product: compound 509: ¹³C-NMR: δ=167.6, 153.5, 147.6,142.8, 141.1, 135.3, 125.3, 123.8, 121.5, 116.4, 106.3, 70.0, 67.0,59.9, 56.2, 55.8, 45.8, 43.8, 40.3, 40.3, 36.4, 28.7, 27.5, 25.7, 25.6,23.3, 22.1, 20.4, 20.0, 18.1, 17.9, 14.1, 12.1, −5.0, −5.1 ppm; Secondeluted product: compound 508: ¹³C-NMR: δ=168.5, 153.5, 148.7, 142.7,138.7, 135.4, 124.9, 123.5, 121.5, 116.4, 106.4, 70.0, 67.0, 60.2, 56.1,55.7, 45.8, 43.8, 40.8, 40.2, 36.4, 28.7, 27.5, 25.7, 25.6, 23.3, 22.1,20.0, 18.1, 17.9, 14.1, 12.4, 12.1, −5.0, −5.1 ppm.

Preparation 10: Compound 512 and 513

Intermediate 2: 203 (1 g, 1.68 mmol).

First eluted product: compound 513: ¹³C-NMR: δ=167.6, 153.4, 148.1,143.0, 140.8, 135.2, 125.2, 123.8, 121.5, 116.2, 106.4, 70.0, 67.0,59.9, 56.6, 56.0, 45.8, 43.8, 40.7, 39.6, 36.4, 28.7, 27.0, 25.7, 25.6,23.2, 21.8, 20.6, 20.5, 18.1, 17.9, 14.1, 12.1, −4.9, −5.1 ppm; Secondeluted product: compound 512: ¹³C-NMR: δ=168.5, 153.4, 149.3, 142.9,138.5, 135.3, 124.9, 123.4, 121.5, 116.3, 106.5, 70.1, 67.0, 60.2, 56.5,56.0, 45.7, 43.8, 41.1, 39.6, 36.4, 28.7, 27.0, 25.7, 25.6, 23.2, 21.8,20.7, 18.0, 17.9, 14.2, 12.4, 12.2, −5.0, −5.1, −5.1 ppm.

General Procedure 3 (Preparations 11 to 14) [5→5′]

To a solution, maintained at about −70° C., of the ester 5 (0.34 mmol)in dry THF (5 ml) was added DIBAL (1M in hexanes, 1 ml, 1 mmol). After30 min at the same temperature, the temperature of the mixture was thenallowed to rise to −20° C. over 1 h, after which recooling at −70° C.was resumed for the addition of methanol (0.5 ml) to quench thereaction. The mixture was then partitioned between water and ethylacetate, and worked up according to the standard procedure above to givethe alcohol 5′.

Preparation 11: Compound 510

Intermediate 5: 508 (0.23 g).

Compound 510: ¹³C-NMR: δ=153.5, 142.9, 141.2, 135.3, 134.5, 125.4,123.3, 121.5, 116.3, 106.4, 70.0, 68.6, 67.0, 56.2, 56.1, 45.7, 43.8,40.4, 40.3, 36.4, 28.7, 27.7, 25.7, 25.6, 23.3, 22.0, 20.4, 18.1, 17.9,13.9, 12.1, −4.9, −5.1 ppm.

Preparation 12: Compound 511

Intermediate 5: 509 (0.18 g, 0.26 mmol).

Compound 511: ¹³C-NMR: δ=153.5, 142.9, 140.8, 135.3, 134.0, 128.3,122.7, 121.5, 116.3, 106.4, 70.1, 67.0, 61.7, 56.2, 56.1, 45.7, 43.8,40.3, 36.4, 28.7, 27.6, 25.7, 25.6, 25.4, 23.3, 22.0, 21.2, 20.3, 18.0,17.9, 12.1, −5.0, −5.1 ppm.

Preparation 13: Compound 514

Intermediate 5: 512 (0.36 g, 0.53 mmol).

Compound 514: ¹³C-NMR: δ=153.4, 143.1, 141.6, 135.1, 134.3, 125.4,123.2, 121.5, 116.2, 106.5, 70.1, 68.6, 67.0, 56.7, 56.1, 45.8, 43.8,40.7, 39.6, 36.4, 28.8, 27.1, 25.7, 25.6, 23.2, 21.9, 21.1, 18.0, 17.9,13.9, 12.1, −5.0, −5.1 ppm.

Preparation 14: Compound 515

Intermediate 5: 513 (0.23 g).

Compound 515: ¹³C-NMR: δ=153.4, 143.1, 141.3, 135.2, 133.9, 128.3,122.6, 121.5, 116.2, 106.5, 70.1, 67.0, 61.7, 56.7, 56.1, 45.8, 43.8,40.6, 39.6, 36.4, 28.8, 27.1, 25.7, 25.6, 23.2, 21.9, 21.2, 21.0, 18.0,17.9, 12.1, −4.9, −5.1 ppm.

General Procedure 4 (Preparation 15) [5→5′]

To a solution, maintained at about −70° C., of the ester 5 (0.6 mmol) indry THF (5 ml) was added the alkyl-lithium. After 1 h at the sametemperature, the reaction was quenched with methanol (0.5 ml), and themixture partitioned between ether and water. Standard work-up gave thealcohol 5.

Preparation 15: Compound 516 [5→5′]

Intermediate 5: 508 (0.4 g).

Alkyl-lithium: Ethyl-lithium (0.8 M in diethyl ether, 2 ml).

Compound 516: ¹³C-NMR: δ=153.5, 143.0, 140.1, 137.3, 135.2, 124.4,123.8, 121.5, 116.3, 106.4, 78.2, 70.1, 67.0, 56.3, 56.2, 45.8, 43.8,40.4, 40.3, 36.4, 31.6, 28.8, 27.6, 25.7, 25.6, 23.3, 22.0, 20.4, 18.1,17.9, 13.3, 12.1, 7.4, −4.9, −5.1 ppm.

Preparation 16: Compound 517 [2→5]

To a solution/suspension, maintained at about 5° C., of compound 204(0.32 g, 0.5 mmol) and methyl-triphenylphosphonium bromide (0.59 g, 1.57mmol) in dry THF (5 ml) was added potassium tert-butoxide (1M solutionin THF, 1.4 ml). After 2 h at the same temperature, the mixture waspartitioned between water and ether, and worked up as standard to giveCompound 517, recrystallised from ether-methanol: ¹³C-NMR: δ=153.5,147.6, 143.0, 136.8, 135.3, 129.6, 121.5, 116.3, 110.3, 106.4, 70.0,67.0, 56.3, 45.8, 43.8, 40.3, 40.2, 36.4, 28.8, 27.5, 25.7, 25.6, 23.3,22.0, 20.4, 18.1, 17.9, 12.6, 12.1, 5.3, 5.2, −5.0, −5.1 ppm.

Preparation 17: Compound 518 [2→5]

Tetrabromomethane (288 mg, 0.87 mmol) was dissolved in dry THF (3.6 ml).

Triphenylphosphine (456 mg 1.74 mmol) was added and the reaction mixturestirred for 30 minutes at room temperature. A solution of Compound 203(261 mg; 0.435 mmol) and triethylamine (0.06 ml; 0.43 mmol) in THF (3.2ml) was added. The reaction mixture was stirred for 90 minutes at roomtemperature, quenched with water (15 ml) and filtered through Decalitefilter aid. The filter was washed with pentane (2×25 ml). The filtratewas extracted with water (3×15 ml) and saturated aqueous sodium chloride(15 ml), dried and concentrated in vacuo. The residue was purified bychromatography (0.5% ether in petroleum ether). 518: ¹³C-NMR: δ 153.4,145.7, 142.8, 137.0, 135.3, 124.6, 121.5, 116.3, 106.5, 88.1, 70.1,67.0, 56.4, 56.0, 45.7, 43.8, 40.8, 39.6, 36.4, 29.5, 28.7, 27.0, 25.7,25.6, 25.4, 23.1, 21.8, 20.5, 18.1, 17.9, 12.1, −5.0, −5.1 ppm.

General Procedure 5 (Preparations 18, 19, 20) [6-7→9→10, including6→7-8]

To a solution, maintained at about −70° C., of the dichloro-intermediate6 (1 mmol) in dry THF (5 ml) was added n-butyl-lithium (1.33 ml, 1.5M inhexanes, 2 molar eq.). The temperature of the mixture was then allowedto rise to 0° C. momentarily to ensure effective conversion to theintermediate lithio-derivative 7. Quenching by partitioning betweensaturated ammonium chloride solution and ether, and work up at thisstage mixture gave intermediate 8. Alternatively recooling of thesolution of 7 at −70° C. was resumed, and addition of a carbonylcompound (1.5 mmol), was made. After 30 minutes at the same temperatureand slow warming up to room temperature, the mixture was partitionedbetween saturated ammonium chloride solution and ether, and worked up asstandard to give compound 9.

To a solution of, maintained at about 5° C., of this intermediatealcohol 9 (ca. 0.4 mmol) in dry dichloromethane (8 ml) was addedMartin's sulfurane (0.54 g, 2 molar equiv.).

After stirring at the same temperature for 1 h, the reaction mixture waspartitioned between ether and 20% sodium hydroxide solution. Standardwork-up gave the Product 10.

Preparation 18: Compound 1001 via Intermediate 701 and Compound 901

Intermediate 6: 601 (0.62 g, 0.97 mmol).

Carbonyl compound: Isobutyraldehyde (0.13 ml).

Compound 901: ¹³C-NMR: δ=153.5, 142.6, 135.4, 121.4, 116.4, 106.4, 90.4,80.3, 70.0, 68.0, 67.0, 55.9, 55.8, 45.6, 43.8, 39.5, 36.4, 34.5, 28.7,27.6, 26.3, 25.7, 25.6, 23.1, 22.0, 21.4, 18.1, 18.0, 17.9, 17.2, 12.4,−5.0, −5.1 ppm.

Intermediate 9: 901 (0.27 g).

The product 10 was further purified by crystallisation fromether-methanol.

Compound 1001: ¹³C-NMR: δ=153.4, 146.0, 142.8, 135.3, 121.5, 116.4,106.5, 105.4, 96.5, 78.9, 70.1, 67.0, 56.1, 56.0, 45.6, 43.8, 39.6,36.4, 28.7, 28.4, 26.4, 25.7, 25.6, 24.4, 23.1, 22.1, 21.6, 20.6, 18.1,17.9, 12.3, −5.0, −5.1, −5.1 ppm.

Preparation 19: Compound 801 via Intermediate 701

Intermediate 6: 601 (0.64 g).

Compound 801: ¹³C-NMR: δ=153.4, 142.6, 135.4, 121.4, 116.5, 106.5, 89.0,70.1, 68.4, 67.0, 55.9, 55.5, 45.6, 43.8, 39.6, 36.4, 28.7, 27.6, 26.4,25.7, 25.6, 23.1, 22.0, 21.2, 18.1, 17.9, 12.1, −4.9, −5.1, −5.1 ppm.

Preparation 20: Compound 802 via Intermediate 702

Intermediate 6: 602 (0.66 g).

Compound 802: ¹³C-NMR: δ=153.4, 142.9, 135.3, 121.5, 116.3, 106.5, 89.1,70.1, 69.0, 67.1, 56.0, 55.4, 45.8, 43.8, 39.5, 36.4, 28.8, 27.3, 25.7,25.6, 23.2, 21.7, 20.8, 18.1, 17.9, 12.1, −4.9, −5.1-5.1 ppm.

1-Bromo-3-hydroxy-3-ethyl-pentyne

To a solution of 3-hydroxy-3-ethyl-1-pentyne (20 mmol) in dry THF (40ml) at room temperature was added n-butyl-lithium (42 mmol, 1.6M inhexanes) during 10 minutes. After stirring for 30 minutes, the solutionwas cooled to −40° C. and a solution of bromine (1.13 ml, 3.52 g, 22mmol) in dry THF (20 ml), also cooled to −40° C., was added, during 20minutes, followed by re-heating to 25° C., during about 1 hour. Standardwork-up after addition of ether and water (chromatography: 0% to 10%ether in petroleum ether) gave the title compound.

1-Bromo-3-hydroxy-3-methyl-1-butyne

When 3-hydroxy-3-methyl-1-butyne was used as starting material in theabove preparation the product was 1-bromo-3-hydroxy-3-methyl-1-butyne.This was also prepared from 3-hydroxy-3-methyl-1-butyne by treatment atroom temperature of an acetone solution with silver nitrate (0.3 eq.)and then, after 20 min, N-bromosuccinimide (1 molar equiv.). After 12 hether was added, the solution filtered, and the filtrate worked-up asstandard to give an oil that was distilled (b.p. 67° C./18 mmHg):¹³C-NMR: δ=31.0, 31.0, 42.6, 66.1, 84.3 ppm.

General Procedure 6 (Preparations 21 to 23) [8→10]

To a solution, maintained at about 25° C., in dry pyrrolidine (5 ml) ofthe Intermediate 8 (0.2 mmol) was added the side chain building block(0.8 mmol; 4 molar equiv.), CuI 4 mg; 0.1 molar equiv.) andbis-triphenylphosphine-palladium dichloride (7 mg; 0.05 molar equiv.).After stirring at the same temperature for 17 h, the reaction mixturewas partitioned between ether and saturated ammonium chloride solutionand worked up as standard to give 10.

Preparation 21: Compound 1003

Intermediate 8: 802 (66 mg, 0.11 mmol).

Side chain building block: 1-Bromo-3-hydroxy-3-methyl-1-butyne (75 mg).

Compound 1003: ¹³C-NMR: δ=153.4, 142.7, 135.4, 121.4, 116.3, 106.5,85.6, 80.4, 70.1, 67.2, 67.0, 65.4, 65.2, 55.8, 55.4, 45.7, 43.8, 39.2,36.4, 31.0, 28.8, 28.0, 27.2, 25.7, 25.6, 23.3, 21.7, 20.3, 18.1, 17.9,12.2, −5.0, −5.1, −5.1 ppm.

Preparation 22: Compound 1005

Intermediate 8: 801 (114 mg).

Side chain building block: 1-bromo-3-hydroxy-3-ethyl-1-pentyne.

Compound 1005: ¹³C-NMR: δ=153.4, 142.4, 135.5, 121.4, 116.5, 106.5,85.0, 78.5, 72.5, 70.1, 69.3, 67.0, 64.9, 55.8, 55.6, 45.7, 43.7, 39.5,36.4, 34.1, 28.6, 28.3, 26.3, 25.7, 25.6, 23.1, 22.0, 20.7, 18.1, 17.9,12.2, 8.3, −5.0, −5.1, −5.1 ppm.

Preparation 23: Compound 1007

Intermediate 8: 802 (118 mg).

Side chain building block: 1-bromo-3-hydroxy-3-ethyl-1-pentyne.

Compound 1007: ¹³C-NMR: δ=153.4, 142.7, 135.4, 121.4, 116.3, 106.5,85.0, 78.8, 72.5, 70.1, 69.2, 67.0, 65.4, 55.8, 55.4, 45.7, 43.8, 39.2,36.4, 34.1, 28.8, 27.9, 27.1, 25.7, 25.6, 23.3, 21.7, 20.2, 18.1, 17.9,12.2, 8.3, −5.0, −5.1, −5.1 ppm.

Preparation 24: Compound 536 [2→5]

Tetrabromomethane (8789 mg, 2.65 mmol) was dissolved in dry THF (9 ml).Triphenylphosphine (1800 mg, 6.8 mmol) was added and the reactionmixture stirred for 30 minutes at room temperature. A solution ofcompound 202 (794 mg, 1.32 mmol) and triethylamine (0.18 ml; 1.4 mmol)in THF (8 ml) was added. The reaction mixture was stirred for 90 minutesat room temperature, quenched with saturated aqueous hydrogen carbonate(15 ml) and filtered through Decalite filter aid. The filter was washedwith pentane (2×25 ml). The filtrate was extracted with water (3×15 ml)and saturated aqueous sodium chloride (15 ml), dried and concentrated invacuo. The residue was purified by chromatography (1.5% ether inpetroleum ether) and crystallised from ether-methanol.

Compound 536: ¹H-NMR: δ=6.86 (d,1H,J=9.9 Hz), 6.45 (d,1H), 6.02(dd,1H,J=9.9 Hz and 15.3 Hz), 5.82 (d,1H), 5.76 (dd,1H, J=8.8 Hz and15.3 Hz), 4.98 (m,1H), 4.94 (m,1H), 4.53 (m,1H), 4.21 (m,1H), 2.87(m,1H), 2.55 (dd,1H), 2.30 (bd,1H), 2.18 (m,1H), 2.10-1.00 (m,13H), 1.07(d,3H), 0.89 (s,9H), 0.86 (s,9H), 0.56 (s,3H), 0.08-0.01 (m,12H) ppm.

Preparation 25: Compound 538 [2→5]

Compound 202 (599 mg, 1 mmol) and bromotrichloromethane (0.095 ml, 1mmol) were dissolved in DCM (30 ml) and cooled to −20 C. Whilemaintaining the temperature between −20 and −10 C a solution oftris(dimethylamino)phosphine (0.40 ml, 2.2 mmol) in DCM (30 ml) wasadded dropwise. After stirring 30 minutes at room temperature pentane(150 ml) and water (30 ml) were added. The organic layer was isolated,washed with water (3×25 ml), and saturated aqueous sodium chloride (25ml), dried, concentrated in vacuo and purified by chromatography (1%ether in petroleum ether).

Compound 538: ¹H-NMR: δ=6.45 (d,1H), 6.34 (d,1H,J=10.3 Hz), 6.10(dd,1H,J=10.3 Hz and 15.0 Hz) 5.82 (d,1H), 5.69 (dd,1H,J=8.8 Hz and 15.0Hz), 4.98 (m,1H), 4.94 (m,1H), 4.53 (m,1H), 4.21 (m,1H), 2.87 (m,1H),2.55 (dd,1H), 2.30 (bd,1H), 2.19 (m,1H), 2.10-1.13 (m,13H), 1.07 (d,3H),0.89 (s,9H), 0.86 (s,9H), 0.56 (s,3H), 0.08-0.02 (m,12H) ppm.

Preparation 26: Compound 540 [2-5]

Cyclobutyltriphenylphosphonium bromide (516 mg, 1.3 mmol) was suspendedin dry THF (4.5 ml) and cooled to −6 C. n-Butyl lithium (1.6 M inhexane, 1.0 ml, 1.6 mmol) was added, the reaction mixture stirred for 20minutes at room temperature, and then again cooled to −6° C. A solutionof Compound 205 (400 mg, 0.65 mmol) andtris(2-(2-methoxyethoxy)ethyl)amine (42 μl, 0.13 mmol) was added and thereaction mixture stirred at room temperature for 3 hours. Water (20 ml)and petroleum ether (100 ml) were added. The organic layer was isolated,washwed with water (3×20 ml) and aqueous saturated sodium chloride (20ml), dried, concentrated in vacuo, and purified by chromatography (0.5%ether in petroleum ether).

Compound 540: ¹H-NMR: δ=6.46 (d,1H), 6.00 (d,1H,J=15.7 Hz), 5.81 (d,1H),5.33 (dd,1H, J=8.8 Hz and 15.7 Hz), 4.98 (t,1H), 4.93 (t,1H), 4.53(m,1H), 4.21 (m,1H), 2.87 (m,1H), 2.73 (m,4H), 2.55 (dd,1H), 2.30(m,1H), 2.22-1.00 (m,16H), 1.56 (bs,3H), 1.05 (d,3H), 0.89 (s,9H), 0.86(s,9H), 0.56 (s,3H), 0.10-0.02 (m,12H) ppm.

Preparation 27: Compound 542

Alkyl-triphenylphosphonium salt: Cyclopentyl-triphenylphosphoniumbromide (1.40 g, 3.4 mmol).

Intermediate 2: 202 (1 g, 1.67 mmol).

Purification of compound 542 by chromatography (2% v:v, ether inpetroleum ether).

Compound 542: ¹H-NMR: δ=6.45 (d,1H), 6.02 (dd,1H,J=11.1 Hz and 14.9 Hz),5.87 (m,1H), 5.81 (d,1H), 5.37 (dd,1H,J=8.8 Hz and 11.1 Hz), 4.98(t,1H), 4.94 (t,1H), 4.53 (m,1H), 4.21 (m,1H), 2.87 (m,1H), 2.55(dd,1H), 2.37-2.24 (m,5H), 2.20-1.86 (m,4H), 1.83-0.80 (m,14H), 1.05(d,3H), 0.90 (s,9H), 0.86 (s,9H), 0.56 (s,3H), 0.08-0.03 (m,12H) ppm.

Preparation 28: Compound 544

Alkyl-triphenylphosphonium salt: Cyclohexyl-triphenylphosphonium bromide(1.44 g, 3.4 mmol).

Intermediate 2: 202 (1 g, 1.67 mmol).

Purification of compound 544 by chromatography (2% v:v, ether inpetroleum ether).

Compound 544: ¹H-NMR: δ=6.45 (d,1H), 6.20 (dd,1H,J=10.7 Hz and 14.9 Hz),5.81 (d,1H), 5.70 (d,1H,J=10.7 Hz), 5.43 (dd,1H,J=8.4 Hz and 14.9 Hz),4.97 (m, H), 4.93 (m,1H), 4.53 (m,1H), 4.21 (m,1H), 2.86 (m,1H), 2.56(dd,1H), 2.35-0.80 (m,25H), 1.04 (d,3H), 0.89 (s,9H), 0.86 (s,9H), 0.56(s,3H), 0.08-0.02 (m,12H) ppm.

Preparation 29: Compound 546

Alkyl-triphenylphosphonium salt: Cyclopropyl-triphenylphosphoniumbromide (766 mg, 2 mmol).

Intermediate 1: Compound 103 (573 g, 1 mmol).

Purification by chromatography (2% v:v, ether in petroleum ether) togive1(S),3(R)-bis(tert-butyldimethylsilyloxy)-20(R)-cyclopropylidenemethyl-9,10-seco-pregna-5(E),7(E),10(19)-triene(compound 546).

Compound 546: ¹³C-NMR: δ=153.4, 143.3, 135.0, 125.0, 121.6, 117.7,116.1, 106.4, 70.1, 67.0, 56.8, 56.0, 45.7, 43.8, 39.7, 39.6, 36.4,28.8, 27.1, 25.7, 25.6, 23.3, 21.9, 20.7, 18.0, 17.9, 12.2, 1.8, 1.7,−4.9, −5.0, −5.1, −5.1 ppm.

Preparation 30: Compound 548

Alkyl-triphenylphosphonium salt: Cyclopropyl-triphenylphosphoniumbromide (606 mg, 1.6 mmol).

Intermediate 1: Compound 101 (458 g, 0.8 mmol).

Purification by chromatography (2% v:v, ether in petroleum ether) togive1(S),3(R)-bis(tert-butyldimethylsilyloxy)-20(S)-cyclopropylidenemethyl-9,10-seco-pregna-5(E),7(E),10(19)-triene(compound 548).

Compound 548: ¹³C-NMR: δ=153.4, 143.2, 135.1, 124.4, 121.6, 118.2,116.2, 106.4, 70.1, 67.0, 56.6, 56.3, 45.7, 43.8, 40.2, 39.6, 36.4,28.8, 27.4, 25.7, 25.6, 23.4, 22.1, 20.3, 18.1, 17.9, 12.0, 2.2, 1.7,−4.9, −5.0, −5.1 ppm.

Preparation 31: Compound 550

To a solution, maintained at about −70° C., of the lithio-derivative 302(prepared from the precursor24-diphenylphosphinoyl-1(S),3(R)-di-TBS-oxy-9,10-seco-chola-5(E),7(E),10(19),22(E)-tetra-ene(0.21 g, 0.27 mmol) and n-butyl-lithium (0.55 ml)) in dry THF (2 ml))was added dropwise a solution of diethyl ketone (0.03 ml) in dry THF(0.3 ml)). After stirring at the same temperature for 30 min, thereaction mixture was kept at room temperature for 2 h, before quenchingwith wet THF, partitioning between ether and water, and standard work-un(chromatography: 1% h v:v ether in petroleum ether) to give the compound550:

¹H-NMR: δ (CDCl₃)=6.45 (d,1H), 6.18 (dd,1H), 5.81 (d,1H), 5.71 (d,1H),5.43 (dd,1H), 4.98 (m,1H), 4.93 (m,1H), 4.53 (m,1H), 4.21 (m,1H), 2.87(d,1H), 2.54 (dd,1H), 2.30 (bd,1H), 2.20-1.10 (m,18H), 1.05 (d,3H), 1.00(m,6H), 0.88 (s,9H), 0.86 (s,9H), 0.56 (s,3H), 0.05 (bs,12H) ppm.

Preparation 32: Compound 303

A solution of24-diphenylphosphinoyl-1(S),3(R)-di-TBS-oxy-9,10-seco-chola-5(E),7(E),10(19),22(E)-tetra-ene(see Preparation 33) (2.4 g, 3.1 mmol) in DCM (5 ml) was briefly cooledon a dry-ice bath for the addition of liquid sulphur dioxide (10 ml).The mixture was stirred under reflux (dry-ice cold finger condenser)without further cooling for 30 min, after which the solvents wereremoved in vacuo and the product crystallised from ether to give themajor isomer sulphur dioxide adduct of24-diphenylphosphinoyl-1(S),3(R)-di-TBS-oxy-9,10-seco-chola-5(E),7(E),10(19),22(E)-tetra-ene:

¹H-NMR: δ (CDCl₃)=7.80-7.65 (m,4H), 7.55-7.40 (m,6H), 5.34 (m,2H), 4.60(m,2H), 4.36 (m,1H), 4.18 (m,1H), 3.92 (m,1H), 3.58 (bd,1H), 3.04(m,2H), 2.56 (m,1H), 2.14 (bd,1H), 2.05-1.00 (m,15H), 0.88 (d,3H), 0.87(bs,18H), 0.55 (s,3H), 0.05 (bs,12H) ppm.

To a stirred solution, maintained at about 5° C., of this adduct (1.8 g,2.1 mmol) in dry DCM (30 ml) was added dropwise boron trifluorideetherate (1.2 ml). After 16 h at 3-5° C., 5% aq. sodium hydrogencarbonate solution was added before diluting with ether and performingthe standard work-up (chromatography: 25% v:v acetone in DCM) to give asthe main product one isomer of the sulphur dioxide adduct of24-diphenylphosphinoyl-3(R)-hydroxy-1(S)-TBS-oxy-9,10-seco-chola-5(E),7(E),10(19),22(E)-tetra-ene:

¹H-NMR: δ (CDCl₃)=7.75-7.67 (m,4H), 7.48-7.42 (m,6H), 5.34 (m,2H), 4.68(m,2H), 4.36 (bt,1H), 4.21 (m,1H), 3.92 (bd,1H), 3.58 (bd,1H), 3.04(m,2H), 2.57 (m,1H), 2.24 (bd,1H), 2.00-0.95 (m,16H), 0.89 (d,3H), 0.87(s,9H), 0.54 (s,3H), 0.08 (s,3H), 0.07 (s,3H) ppm.

A stirred solution, maintained at about 5° C., of this alcohol (0.61 g,0.84 mmol) in dry DCM (10 ml) was treated with the Dess-Martinperiodinane (0.5 g). After 30 min 5% aq. sodium hydrogen carbonatesolution and 10% aq. sodium thiosulphate solution were added andstirring continued for 10 min before dilution with ether and performingthe standard work-up (chromatography: 25% v:v acetone in DCM) to giveone isomer of the sulphur dioxide adduct of24-diphenylphosphinoyl-3-oxo-1(S)-TBS-oxy-9,10-seco-chola-5(E),7(E),10(19),22(E)-tetra-ene.This ketone (0.53 g, 0.72 mmol) was dissolved in THF-methanol (2+5 ml)and treated with sodium borohydride (43 mg) while stirring on an icebath. After the reduction was complete (15 min), the reaction mixturewas partitioned between ethyl acetate and water. Standard work-up(without chromatography) gave the 3(S)-hydroxy compound in admixturewith a small amount of the 3(R)-hydroxy compound already described. Theproduct was heated under reflux in toluene (10 ml)/water (10 ml)containing suspended sodium hydrogen carbonate (1.2 g) for 1 h. Aftercooling, partitioning between ether and water followed by standardwork-up (without chromatography) gave24-diphenylphosphinoyl-3(S)-hydroxy-1(S)-TBS-oxy-9,10-seco-chola-5(E),7(E),10(19),22(E)-tetra-enein admixture with a small amount of the 3(R)-hydroxy compound. Theproduct was submitted to the process of General Procedure 7 (withoutchromatography), using 9-acetyl-anthracene (0.02 g) and DCM to give the5(Z)-isomer. To a stirred, ice-cooled solution of this product in dryDCM (10 ml) and 2,6-lutidine (0.2 ml) was added TBS triflate (0.3 ml),and after allowing to react for 1 h, the mixture was partitioned betweenwater and ether, and worked up as standard (chromatography: 25% v:vacetone in DCM) to give24-diphenylphosphinoyl-1(S),3(S)-Di-TBS-oxy-9,10-seco-chola-5(E),7(E),10(19),22(E)-tetra-ene:

¹H-NMR: δ (CDCl₃)=7.80-7.65 (m,4H), 7.50-7.40 (m,6H), 6.23 (d,1H), 5.94(d,1H), 5.37 (bt,1H), 5.34 (m,2H), 4.91 (bt,1H), 3.93 (m,1H), 3.68(m,1H), 3.05 (m,2H), 2.78 (dd,1H), 2.41 (bd,1H), 2.25-0.90 (m,15H), 0.93(s,9H), 0.87 (s,9H), 0.86 (d,3H), 0.46 (s,3H), 0.10 (s,3H), 0.07 (s,3H),0.06 (s,3H), 0.05 (s,3H) ppm. The lithio-derivative 303 was prepared asrequired from this precursor by adding dropwise n-butyl-lithium (1.6M inhexanes, 1 molar equiv.) to a stirred solution, maintained at about −70°C., in dry THF and keeping at the same temperature for 15 min.

Preparation 33: Compound 302

To a stirred solution of1(S),3(R)-di-TBS-oxy-9,10-seco-chola-5(E),7(E),10(19),22(E)-tetra-ene-24-ol(1.17 g, 1.94 mmol) in dry THF (15 ml) at room temperature was addedtriphosgene (0.43 g) and then dropwise pyridine (0.5 ml). After 90 minat the same temperature, the mixture was partitioned between water andether, and worked up as standard (chromatography: 1% v:v ether inpetroleum ether) to give24-chloro-1(S),3(R)-di-TBS-oxy-9,10-seco-chola-5(E),7(E),10(19),22(E)-tetra-ene:

¹³C-NMR: δ (CDCl₃)=153.5, 142.8, 141.8, 135.4, 123.5, 121.5, 116.4,106.4, 70.0, 67.0, 56.2, 55.7, 45.7, 45.5, 43.8, 40.2, 39.6, 36.4, 28.7,27.4, 25.7, 25.6, 23.3, 22.0, 19.9, 18.1, 17.9, 12.1, −5.0, −5.1 ppm.

To a stirred solution, maintained at about −70° C., of this chloride(0.81 g) in dry THF (7 ml) was added dropwise a solution of lithiumdiphenylphosphide (0.4M in dry THF, 3.5 ml). A few drops of water werethen added before warming to room temperature and concentration invacuo, to give an oil. This was taken up in DCM (15 ml) and stirredvigorously with 5% hydrogen peroxide (aq., 12 ml) for 30 min. Themixture was partitioned between water and ether, and worked up asstandard (chromatography: 66% v:v ethyl acetate in petroleum ether) togive24-diphenylphosphinoyl-1(S),3(R)-di-TBS-oxy-9,10-seco-chola-5(E),7(E),10(19),22(E)-tetra-ene:

¹H-NMR: δ (CDCl₃)=7.8-7.6 (m,4H), 7.5-7.4 (m,6H), 6.42 (d,1H), 5.77(d,1H), 5.45-5.25 (m,2H), 4.96 (bs,1H), 4.92 (bs,1H), 4.52 (m,1H), 4.20(m,1H), 3.07 (d,1H), 3.02 (d,1H), 2.84 (bd,1H), 2.55 (dd,1H), 2.27(bd,1H), 2.05-0.90 (m,14H), 0.88 (s,9H), 0.87 (d,3H), 0.86 (s,9H), 0.47(s,3H), 0.05 (s,12H) ppm.

The lithio-derivative 302 was prepared as required from this precursorby adding dropwise n-butyl-lithium (1.6M in hexanes, 1 molar equiv.) toa stirred solution, maintained at about −70° C., in dry THF and keepingat the same temperature for 15 min.

Preparation 34: Compound 534

To a solution, maintained at about −70° C., of the lithio-derivative 302(prepared from the precursor24-diphenylphosphinoyl-1(S),3(R)-di-TBS-oxy-9,10-seco-chola-5(E),7(E),10(19),22(E)-tetra-ene(0.71 g, 0.88 mmol) and n-butyl-lithium (0.55 ml)) in dry THF (8 ml))was added dropwise a solution of hexafluoro-acetone (1 ml of ca. 1Msolution in dry THF). After stirring at the same temperature for 30 min,the reaction mixture was kept at room temperature for 4 h beforequenching with wet THF, partitioning between ether and water, andstandard work-up (chromatography: 5% v:v ether in petroleum ether) togive the compound 534:

¹³C-NMR: δ (CDCl₃)=157.3, 153.5, 142.3, 142.2, 135.6, 121.5, 121.4,121.3, 120.5, 116.6, 106.5, 70.0, 67.0, 56.0, 55.3, 46.0, 43.8, 40.9,40.2, 36.4, 28.7, 27.4, 25.7, 25.6, 23.2, 22.1, 19.4, 18.1, 17.9, 12.1,−4.9, −5.0, −5.1, −5.1 ppm.

Preparation 35: Compound 205 [1→2]

A mixture of compound 101 (5.00 g, 8,7 mmol),1-(triphenylphosphoranylidene)-2-propanone (8.32 g, 26,1 mmol), andtoluene (100 ml) was stirred for 18 hours at 110° C. The reactionmixture was cooled to room temperature, filtered, and concentrated invacuo. The residue was dissolved in pentane (75 ml), stirred for 45minutes at room temperature, and filtered through Decalite filter aid.The filtrate was concentrated in vacuo and purified by chromatography(0-5% v:v ether in petroleum ether).

Compound 205: ¹H-NMR: δ=6.66 (dd,1H,J=8.8 Hz and 16.1 Hz), 6.44 (d,1H),6.00 (d,1H,J=16.1 Hz), 5.81 (d,1H), 4.98 (m,1H),4.93 (m,1H), 4.53(m,1H), 4.21 (m,1H), 2.88 (m,1H), 2.54 (dd,1H), 2.38-2.16 (m,2H), 2.22(s,3H), 2.10-1.85 (m,3H), 1.82-1.18 (m,10H), 1.11 (d,3H), 0.89 (s,9H),0.86 (s,9H), 0.57 (s,3H), 0.08-0.02 (m,12H).

General Procedure 7 (Preparations 101-126, 128) [a→b (B=CH₂)]

A solution of the 5E-Vitamin D intermediate of type a (0.1 mmol), atriplet-sensitizer (0.01 g), and triethylamine (0.05 ml) in a solvent (5ml) in a Pyrex flask was irradiated with light from a high pressureultraviolet lamp, type TQ180Z2 (Hanau) at about 20° C. for 30 minutes(the time was scaled-up proportionally according to the amount ofintermediate a). The reaction mixture (after filtering when anthracenewas used) was partially concentrated in vacuo and purified bychromatography to give the product intermediate of type b.

Preparation 101: Compound 519

Intermediate of type a: 501 (1.29 g, 2.06 mmol).

Sensitizer: Anthracene (0.65 g).

Solvent: DCM (100 ml).

Irradiation: A TQ718Z2 lamp was used for 35 min.

Compound 519: ¹H-NMR: δ=6.23 (d, 1H), 6.13 (dd, 1H, J=11.1 Hz and 14.9Hz), 6.00 (d, 1H), 5.75 (d, 1H, J=11.1 Hz), 5.40 (dd, 1H, J=8.8 Hz and14.9 Hz), 5.17 (m, 1H), 4.85 (m, 1H), 4.36 (m, 1H), 4.18 (m, 1H), 2.82(m, 1H), 2.44 (dd, 1H), 2.21 (dd, 1H), 2.11 (m, 1H), 1.74 (bs, 3H), 1.73(bs, 3H), 2.03-1.14 (m, 13H), 1.04 (d, 3H), 0.87 (s, 18H), 0.55 (s, 3H),0.05 (m, 12H) ppm.

Preparation 102: Compound 521

Intermediate of type a: 502 (63 mg).

Sensitizer: 9-acetyl-anthracene.

Solvent: DCM.

Compound 521: ¹H-NMR in agreement with structure.

Preparation 103: Compound 523

Intermediate of type a: 503 (0.38 g).

Sensitizer: Anthracene (0.32 g).

Solvent: DCM.

Purification of compound 523 by chromatography (0.5% v:v, ether inpetroleum ether).

Compound 523: ¹H-NMR: δ=6.22 (d, J=11.2 Hz, 1H), 6.11 (dd, J=14.9 Hz,J=10.7 Hz, 1H), 6.00 (d, J=11.2 Hz, 1H), 5.76 (d, J=10.7 Hz; 1H), 5.41(dd, J=9.6 Hz, J=14.9 Hz, 1H), 5.17 (m, 1H), 4.86 (m, 1H), 4.36 (m, 1H),4.18 (m, 1H), 2.80 (m, 1H), 2.43 (dd, J=3.8 Hz, J=13.0 Hz, 1H), 2.20(dd, J=7.5 Hz, J=13.0 Hz, 1H), 2.15-1.0 (m, 14H), 1.75 (bs, 3H), 1.72(bs, 3H), 0.93 (d, J=6.5 Hz, 3H), 0.87 (s, 9H), 0.86 (s, 9H), 0.49 (s,3H), 0.05 (bs, 12H) ppm.

Preparation 104: Compound 524

Intermediate of type a: 504 (61 mg).

Sensitizer: Anthracene (0.05 g).

Solvent: DCM.

Purification of compound 524 by chromatography (0-2% v:v, ether inpetroleum ether).

Compound 524: ¹H-NMR: δ=6.33 (bd, J=10.3 Hz, 1H), 6.22 (d, J=11.2 Hz,1H), 6.11 (dd, J=10.3 Hz, J=15.3 Hz, 1H), 6.01 (d, J=11.2 Hz,1H), 5.55(dd, J=15.3 Hz, J=8.8 Hz, 1H), 5.17 (bs, 1H), 4.86 (bs, 1H), 4.18 (m,1H), 2.81 (m, 1H), 2.44 (m, 1H), 2.25-1.0 (m, 20H), 1.06 (d, J=6.8 Hz,3H), 0.87 (s, 18H), 0.55 (s, 3H), 0.05 (bs, 12H) ppm.

Preparation 105: Compound 525

Intermediate of type a: 505 (0.24 g).

Sensitizer: Anthracene (0.23 g).

Solvent: DCM.

Purification of compound 525 by chromatography (0-2% v:v, ether inpetroleum ether).

Compound 525: ¹H-NMR: δ=6.35 (bd, J=10.3 Hz, 1H), 6.22 (bd, J=11.5 Hz,1H), 6.08 (dd, J=10.3 Hz, J=15.4 Hz, 1H), 6.00 (d, J=11.5 Hz, 1H), 5.55(dd, J=9.4 Hz, J=15.4 Hz, 1H), 5.17(bd, 1H), 4.86 (bd, 1H), 4.36 (m,1H), 4.18 (m, 1H), 2.80 (bd, J=12.2 Hz, 1H), 2.43 (dd, J=3.6, J=13.0 Hz,1H), 2.20 (dd, J=7.3 Hz, J=13.0 Hz, 1H), 2.15-1.0 (m, 18H), 0.97 (d,J=6.8 Hz, 3H), 0.87 (s, 9H), 0.87 (s, 9H), 0.50 (s, 3H), 0.05 (bs, 12H)ppm.

Preparation 106: Compound 526

Intermediate of type a: 506 (0.4 g).

Sensitizer: Anthracene (0.34 g).

Solvent: DCM.

Purification of compound 526 by chromatography (0.5% v:v, ether inpetroleum ether).

Compound 526: ¹H-NMR: δ=6.22 (d, J=11.2 Hz, 1H), 6.00 (d, J=11.2 Hz,1H), 5.84 (dd, J=15 Hz, J=10.7 Hz, 1H), 5.65 (m, 1H), 5.34 (dd, J=15 Hz,J=8.7 Hz, 1H), 5.17 (bs, 1H), 4.85 (bs, 1H), 4.36 (m,1H), 4.18 (m, 1H),2.69 (m, 5H), 2.44 (m, 1H), 2.25-1.10 (m, 17H), 1.03 (d, J=6.7 Hz, 3H),0.87 (s, 18H), 0.54 (s, 3H), 0.05 (bs, 12H) ppm.

Preparation 107: Compound 527

Intermediate of type a: 507 (0.38 g).

Sensitizer: Anthracene (0.32 g).

Solvent: DCM.

Purification of compound 527 by chromatography (0.5% v:v, ether inpetroleum ether).

Compound 527: ¹³C-NMR: δ=148.2, 142.3, 140.9, 137.5, 134.7, 123.5,123.0, 120.9, 117.6, 110.9, 71.9, 67.3, 56.7, 56.0, 45.8, 45.7, 44.6,40.6, 39.7, 31.0, 29.7, 28.7, 27.1, 25.7, 25.6, 23.2, 21.8, 21.3, 18.0,17.9, 16.9, 12.0, −4.9, −5.0, −5.3 ppm.

Preparation 108: Compound 528

Intermediate of type a: 510 (0.15 g).

Sensitizer: 9-acetyl-anthracene (0.02 g).

Solvent: DCM.

Compound 528: ¹H-NMR in agreement with structure.

Preparation 109: Compound 529

Intermediate of type a: 511 (24 mg, 0.04 mmol).

Sensitizer: 9-acetyl-anthracene.

Solvent: Toluene.

Compound 529: ¹H-NMR in agreement with structure.

Preparation 110: Compound 530

Intermediate of type a: 514 (125 mg, 0.2 mmol).

Sensitizer: 9-acetyl-anthracene.

Solvent: DCM.

Compound 530: ¹H-NMR in agreement with structure.

Preparation 111: Compound 531

Intermediate of type a: 515 (17 mg).

Sensitizer: Anthracene (0.01 g).

Solvent: Toluene (2 ml).

Compound 531: ¹H-NMR in agreement with structure.

Preparation 112: Compound 532

Intermediate of type a: 516 (90 mg).

Sensitizer: 9-acetyl-anthracene.

Solvent: DCM.

Compound 532: ¹H-NMR in agreement with structure.

Preparation 113: Compound 533

Intermediate of type a: 517 (0.10 g, 0.16 mmol).

Sensitizer: 9-acetyl-anthracene.

Solvent: DCM.

Compound 533: ¹H-NMR: δ=6.23 (d, 1H), 6.03 (d,1H, J=15.6 Hz), 6.01 (d,1H), 5.88 (dd, 1H, J=8.4 Hz and 15.6 Hz), 5.17 (m, 1H), 4.86 (m, 1H),4.78 (m, 1H), 4.72 (m, 1H), 4.37 (m, 1H), 4.18 (m, 1H), 2.83 (m, 1H),2.45 (dd,1H), 2.30-2.06 (m, 2H), 2.05-1.95 (m, 2H), 1.07 (d, 3H), 0.87(s, 18H), 1.93-0.80 (m, 12H), 0.67 (m, 2H), 0.56 (s, 3H), 0.41 (m, 2H),0.05 (m, 6H), 0.04 (m, 6H) ppm.

Preparation 114: Compound 520

Intermediate of type a: 518 (57 mg).

Sensitizer: Anthracene (0.05 g).

Solvent: DCM.

Purification of compound 520 by chromatography (0.5% v:v, ether inpetroleum ether).

Compound 520: ¹H-NMR: δ=6.87 (d, J=9.9 Hz, 1H), 6.22 (d, J=11.2 Hz, 1H),6.00 (d, J=11.2 Hz, 1H), 6.01 (dd, J=9.9 Hz, J=15.3 Hz, 1H), 5.81 (dd,J=15.3 Hz, J=9.5 Hz, 1H), 5.17 (bm, 1H), 4.85 (bm, 1H), 4.36 (m, 1H),4.19 (m, 1H), 2.81 (bm, 1H), 2.44 (dd, 1H), 2.2-1.0 (m, 15H), 0.95 (d,J=6.6 Hz, 3H), 0.87 (bs, 18H), 0.50 (s, 3H), 0.05 (bs, 12H) ppm.

Preparation 115: Compound 1002

Intermediate of type a: 1001 (104 mg, 0.17 mmol).

Sensitizer: 9-acetylanthracene.

Solvent: tert-butyl methyl ether.

Compound 1002: ¹H-NMR in agreement with structure.

Preparation 116: Compound 1004

Intermediate of type a: 1003 (41 mg).

Sensitizer: 9-acetylanthracene (10 mg).

Solvent: Toluene (4 ml).

Compound 1004: ¹H-NMR: δ=6.23 (d, 1H), 6.00 (d, 1H), 5.17 (m, 1H), 4.85(m, 1H), 4.37 (m, 1H), 4.18 (m, 1H), 2.82 (m, 1H), 2.44 (dd,1H), 1.52(s, 6H), 1.17 (d, 3H), 0.87 (s, 18H), 2.38-0.80 (m, 16H), 0.56 (s, 3H),0.05 (s, 6H), 0.04 (s, 6H) ppm.

Preparation 117: Compound 1006

Intermediate of type a: 1005 (70 mg, 0.1 mmol).

Sensitizer: Anthracene (80 mg).

Solvent: DCM.

Compound 1006: ¹³C-NMR: δ=148.1, 140.0, 135.2, 122.8, 118.0, 111.0,85.1, 78.4, 72.5, 71.7, 69.3, 67.3, 64.8, 55.7, 55.5, 45.8, 45.5, 44.6,39.5, 34.1, 28.6, 28.3, 26.4, 25.6, 25.6, 23.0, 21.9, 20.7, 18.0, 17.9,12.2, 8.3, −4.9, −5.0, −5.3 ppm.

Preparation 118: Compound 1008

Intermediate of type a: 1007 (68 mg, 0.1 mmol)

Sensitizer: 9-acetylanthracene

Solvent: DCM

Compound 1008: ¹H-NMR: δ=6.23 (d, 1H), 6.00 (d,1H), 5.17 (m, 1H), 4.85(m, 1H), 4.37 (m, 1H), 4.18 (m, 1H), 2.83 (m, 1H), 2.44 (dd,1H), 1.17(d, 3H), 1.03 (t, 6H), 0.87 (s, 18H), 2.38-0.80 (m, 20H), 0.56 (s, 3H),0.05 (m, 6H), 0.04 (m, 6H) ppm.

Preparation 119: Compound 537

Intermediate of type a: 536 (150 mg).

Sensitizer: Anthracene (0.112 g).

Solvent: DCM.

Purification of compound 537 by chromatography (0-2% v:v, ether inpetroleum ether).

Compound 537: ¹H-NMR in agreement with structure.

Preparation 120: Compound 539

Intermediate of type a: 538 (76 mg).

Sensitizer: Anthracene (60 mg).

Solvent: DCM.

Purification of compound 539 by chromatography (0-2% v:v, ether inpetroleum ether).

Compound 539: ¹H-NMR: δ=6.34 (d,1H,J=10.3 Hz), 6.22 (d,1H), 6.09(dd,1H,J=10.3 Hz and 14.9 Hz), 6.01 (d,1H), 5.69 (dd,1H,J=8.8 Hz and14.9 Hz), 5.17 (m,1H), 4.85 (d,1H), 4.36 (m,1H), 4.18 (m,1H), 2.82(m,1H), 2.44 (dd,1H), 2.25-2.10 (m,2H), 2.05-1.10 (m,13H), 1.06 (d,3H),0.87 (s,9H), 0.87 (s,9H), 0.55 (s,3H), 0.07-0.03 (m,12H ppm.

Preparation 121: Compound 541

Intermediate of type a: 540 (120 mg).

Sensitizer: Anthracene (115 mg). Solvent: DCM.

Purification of compound 541 by chromatography (0-2% v:v, ether inpetroleum ether).

Compound 541: ¹H-NMR: δ=6.23 (d,1H), 6.00 (d,1H), 5.99 (d,1H, J=15.6Hz), 5.33 (dd,1H,J=8.8 Hz and 15.6 Hz), 5.17 (m,1H), 4.85 (bd,1H), 4.36(m,1H), 4.18 (m,1H), 2.87-2.60 (m,5H), 2.44 (dd,1H), 2.21 (dd,1H),2.16-1.10 (m,16H), 1.55 (bs,3H), 1.04 (d,3H), 0.87 (bs,18H), 0.55(s,3H), 0.07-0.03 (m,12H) ppm.

Preparation 122: Compound 543

Intermediate of type a: 542 (300 mg).

Sensitizer: Anthracene (200 mg).

Solvent: DCM.

Purification of compound 543 by chromatography (0-2% v:v, ether inpetroleum ether).

Compound 543: ¹H-NMR: δ=6.23 (d,1H), 6.01 (d,1H), 6.01 (dd,1H,J=11.1 Hzand 14.9 Hz), 5.86 (m,1H), 5.36 (dd,1H,J=8.8 Hz and 14.9 Hz), 5.16(m,1H), 4.85 (d,1H), 4.36 (m,1H), 4.18 (m,1H), 2.82 (m,1H), 2.44(dd,1H), 2.37-0.80 (m,23H), 1.04 (d,3H), 0.87 (s,18H), 0.55 (s,3H),0.07-0.03 (m,12H) ppm.

Preparation 123: Compound 545

Intermediate of type a: 544 (214 mg).

Sensitizer: Anthracene (150 mg).

Solvent: DCM.

Purification of compound 543 by chromatography (0-2% v:v, ether inpetroleum ether).

Compound 545: ¹H-NMR: δ=6.23 (d,1H), 6.19 (dd,1H,J=10.7 Hz and 14.9 Hz),6.00 (d,1H), 5.69 (d,1H, J=10.7 Hz), 5.43 (dd,1H,J=8.4 Hz and 14.9 Hz),5.17 (m,1H), 4.85 (d,1H), 4.36 (m,1H), 4.18 (m,1H), 2.81 (m,1H), 2.44(dd,1H), 2.32-0.80 (m,25H), 1.04 (d,3H), 0.87 (s,18H), 0.54 (s,3H),0.07-0.03 (m,12H) ppm.

Preparation 124: Compound 547

Intermediate of type a: 546 (217 mg).

Sensitizer: Anthracene (214 mg).

Solvent: DCM.

Purification by chromatography (0-2% v:v, ether in petroleum ether) togive1(S),3(R)-bis(tert-butyldimethylsilyloxy)-20(R)-cyclopropylidenemethyl-9,10-seco-pregna-5(Z),7(E),10(19)-triene(compound 547).

Compound 547: ¹H-NMR: δ=6.22 (d,1H), 6.00 (d,1H), 5.62 (m,1H), 5.17(m,1H), 4.86 (bd,1H), 4.36 (m,1H), 4.18 (m,1H), 2.79 (m,1H), 2.44(dd,1H), 2.28 (m,1H), 2.20 (dd,1H), 2.02-0.80 (m,17H), 0.96 (d,3H), 0.87(s,9H), 0.87 (s,9H), 0.45 (s,3H), 0.09-0.02 (m,12H) ppm.

Preparation 125: Compound 549

Intermediate of type a: 546 (175 mg).

Sensitizer: Anthracene (157 mg).

Solvent: DCM.

Purification by chromatography (0-2% v:v, ether in petroleum ether) togive1(S),3(R)-bis(tert-butyldimethylsilyloxy)-20(S)-cyclopropylidenemethyl-9,10-seco-pregna-5(Z),7(E),10(19)-triene(compound 549).

Compound 549: ¹H-NMR: δ=6.23 (d,1H), 6.01 (d,1H), 5.61 (m,1H), 5.17(m,1H), 4.86 (bd,1H), 4.37 (m,2H), 4.19 (m,1H), 2.82 (m,1H), 2.44(dd,1H), 2.32 (m,1H), 2.21 (dd,1H), 2.06-0.80 (m,17H), 1.07 (d,3H), 0.87(s,9H), 0.87 (s,9H), 0.56 (s,3H), 0.07-0.03 (m,12H) ppm.

Preparation 126: Compound 551

Intermediate of type a: 550 (0.07 g).

Sensitizer: 9-acetyl-anthracene (0.01 g).

Solvent: DCM (7 ml).

Compound 551: ¹H-NMR (CDCl₃): δ=6.22 (d,1H), 6.17 (dd,1H), 6.00 (d,1H),5.70 (d,1H), 5.43 (dd,1H), 5.16 (bd,1H), 4.85 (bd,1H), 4.36 (m,1H), 4.18(m,1H), 2.79 (dd,1H), 2.43 (dd,1H), 2.25-1.15 (m,19H), 1.04 (d,3H), 1.00(m,6H), 0.86 (bs,18H), 0.54 (s,3H), 0.05 (bs,12H) ppm.

Preparation 127: Compound 552

To a solution, maintained at about −70° C., of the lithio-derivative 303(prepared from the precursor24-diphenylphosphinoyl-1(S),3(S)-Di-TBS-oxy-9,10-seco-chola-5(E),7(E),10(19),22(E)-tetra-ene(0.13 g, 0.16 mmol) and n-butyl-lithium (0.1 ml)) in dry THF (2 ml)) wasadded dropwise a solution of diethyl ketone (0.025 ml) in dry THF (0.25ml)). After stirring at the same temperature for 30 min, the reactionmixture was kept at room temperature for 2 h and then at 40° C. for 1 h,before quenching with wet THF, partitioning between ether and water, andstandard work-up (chromatography: 20% v:v toluene in petroleum ether) togive the compound 552:

¹H-NMR in agreement with structure.

Preparation 128: Compound 535

Intermediate of type a: 534 (0.09 g).

Sensitizer: 9-acetyl-anthracene (0.01 g).

Solvent: DCM (7 ml).

535: ¹H-NMR (CDCl₃): δ=6.96 (d,1H), 6.43 (t,1H), 6.22 (d,1H), 6.19(dd,1H), 6.01 (d,1H), 5.15 (m,1H), 4.84 (d,1H), 4.36 (m,1H), 4.18(m,1H), 2.82 (d,1H), 2.41 (dd,1H), 2.40-2.15 (m,2H), 2.0-1.15 (m,13H),1.15 (d,3H), 0.87 (s,9H), 0.86 (s,9H), 0.57 (s,3H), 0.05 (s,6H), 0.04(s,6H) ppm.

General Procedure 8 (Examples 4, 14-18, 23, 25-27, 29, and 36)

To a mixture, maintained at about 25° C., of an ethyl acetate solution(about 0.3 ml) of the appropriate silyl-protected intermediate (ca. 0.1mmol) in acetonitrile (4 ml) was added (via plastic pipette) 40% aqueoushydrofluoric acid (0.5 ml, ca. 10 mmol). After vigorous stirring at thesame temperature for 1 h, the reaction mixture was partitioned betweenethyl acetate and 3N sodium hydroxide (to alkaline reaction to pH paper)solution before standard work-up to give the compound I.

General Procedure 9 (Examples 1-3, 5-13, 28, 30-33)

To a solution of the appropriate silyl-protected intermediate (0.1 mmol)in THF (4 ml) was added solid TBA-fluoride trihydrate (0.13 g, ca. 0.4mmol), and the solution was heated at 60° C. for one hour. After cooling(and partial concentration in vacuo when the volume of THF in ascaled-up procedure exceeded 30 ml), 0.2 M aqueous sodium hydrogencarbonate solution and ethyl acetate were added. Standard work-up gavethe compound I.

Example 11(S),3(R)-Dihydroxy-9,10-secocholesta-5(Z),7(E),10(19),22(E),24-penta-ene(Compound 1)

A solution of 519 (2.1 g, 3.36 mmol) and tetra-n-butylammonium fluoridetrihydrate (5.42 g, 17.2 mmol) in tetrahydrofuran (70 ml) was heated at60° C. in an argon atmosphere for 60 minutes. After cooling andconcentration in vacuo, the reaction mixture was partitioned betweenethyl acetate and aqueous sodium bicarbonate. The organic phase waswashed with water and brine, dried and concentrated. The residue waspurified by chromatography (50% ethyl acetate in petroleum ether).Fractions containing the title compound were concentrated in vacuo toyield an oil which gave colourless crystals from methyl formate.Compound 1: ¹³C-NMR: δ=147.7, 143.1, 138.2, 132.9, 132.8, 125.3, 125.0,124.3, 117.1, 111.8, 70.8, 66.9, 56.4, 45.9, 45.3, 42.9, 40.5, 40.4,29.1, 27.8, 25.9, 23.6, 22.3, 20.8, 18.2, 12.3 ppm.

Example 21(S),3(R)-Dihydroxy-9,10-secocholesta-5(Z),7(E),10(19),22(Z),24-penta-ene(Compound 2)

Silyl-protected intermediate: 521 (32 mg, 0.05 mmol).

Compound 2: ¹³C-NMR: δ=147.7, 143.1, 136.0, 134.8, 132.9, 125.0, 121.9,120.6, 117.1, 111.7, 70.8, 66.9, 56.6, 56.4, 45.9, 45.3, 42.9, 40.5,34.7, 29.1, 27.4, 26.4, 23.6, 22.3, 20.8, 18.1, 12.4 ppm.

Example 3 1(S),3(R)-Dihydroxy-9,10-seco-20(S)-cholesta-5(Z),7(E),10(19),22(E),24-penta-ene (Compound 3)

Silyl-protected intermediate: 523 (135 mg, 0.22 mmol).

Purification of compound 3 by chromatography (40% v:v, ethyl acetate inpetroleum ether).

Compound 3: ¹³C-NMR: δ=147.7, 143.4, 138.6, 132.8, 132.5, 125.3, 125.0,124.3, 116.9, 111.8, 70.9, 66.9, 56.9, 56.2, 46.0, 45.3, 42.9, 40.9,39.7, 29.1, 27.3, 25.9, 23.5, 22.1, 21.5, 18.3, 12.2 ppm.

Example 41(S),3(R)-Dihydroxy-9,10-seco-26,27-cyclo-cholesta-5(Z),7(E),10(19),22(E),24-penta-ene(Compound 4)

Silyl-protected intermediate: 524 (36 mg).

Purification of compound 4 by chromatography (50% v:v, ethyl acetate inpetroleum ether).

Compound 4: ¹H-NMR: δ=6.37 (d, J=11 Hz, 1H), 6.34 (bd, J=10 Hz, 1H),6.11 (dd, J=10 Hz, J=15 Hz, 1H), 6.01 (d, J=11 Hz, 1H), 5.56 (dd, J=9Hz, J=15 Hz, 1H), 5.32 (bs, 1H), 5.00 (bs, 1H), 4.43 (m, 1H), 4.23 (m,1H), 2.83 (dd, J=4 Hz, J=11 Hz, 1H), 2.59 (dd, J=4 Hz, J=13 Hz, 1H),2.31 (dd, J=6 Hz, J=13 Hz, 1H), 2.2-1.2 (m, 16H), 1.10 (m, 4H), 1.07 (d,J=7 Hz, 3H), 0.58 (s, 3H) ppm.

Example 51(S),3(R)-Dihydroxy-9,10-seco-26,27-cyclo-20(S)-cholesta-5(Z),7(E),10(19),22(E),24-penta-ene(Compound 5)

Silyl-protected intermediate: 525 (165 mg, 0.26 mmol).

Purification of compound 5 by chromatography (50% v:v, ethyl acetate inpetroleum ether).

Compound 5: ¹³C-NMR: δ=147.7, 143.2, 139.5, 132.9, 127.1, 125.0, 123.9,119.1, 117.0, 111.8, 70.8, 66.8, 56.9, 56.2, 46.0, 45.3, 42.9, 40.7,39.8, 29.1, 27.3, 23.5, 22.1, 21.3, 12.3, 2.4, 2.2 ppm.

Example 61(S),3(R)-Dihydroxy-9,10-seco-26,27-methano-cholesta-5(Z),7(E),10(19),22(E),24-penta-ene(Compound 6)

Silyl-protected intermediate: 526 (348 mg, 0.55 mmol).

Isolation from the crude product by direct crystallisation from methylformate, omitting the chromatography step.

Compound 6: ¹³C-NMR: δ=147.6, 143.1, 142.8, 137.2, 133.0, 125.0, 123.9,121.0, 117.1, 111.8, 70.8, 66.9, 56.4, 56.4, 45.9, 45.3, 42.9, 40.4,40.4, 31.3, 29.9, 29.1, 27.8, 23.6, 22.2, 20.8, 17.2, 12.3 ppm.

Example 71(S),3(R)-Dihydroxy-9,10-seco-26,27-methano-20(S)-cholesta-5(Z),7(E),10(19),22(E),24-penta-ene(Compound 7)

Silyl-protected intermediate: 527 (150 mg, 0.23 mmol).

Purification of compound 7 by chromatography (40% v:v, ethyl acetate inpetroleum ether).

Compound 7: ¹³C-NMR: δ=147.7, 143.3, 142.6, 137.6, 132.8, 125.0, 123.8,121.1, 116.9, 111.8, 70.9, 66.9, 56.9, 56.2, 46.0, 45.3, 42.9, 40.8,39.8, 31.3, 29.9, 29.1, 27.3, 23.5, 22.1, 21.5, 17.1, 12.2 ppm.

Example 81(S),3(R),26-Trihydroxy-9,10-secocholesta-5(Z),7(E),10(19),22(E),24(E)-penta-ene(Compound 9)

Silyl-protected intermediate: 528 (148 mg, 0.23 mmol).

Purification of compound 9 by crystallisation from methyl formate.

Compound 9: ¹H-NMR: (in hexadeuterioacetone) δ=6.29 (d, 1H), 6.25 (dd,1H, J=11.1 Hz and 15.1 Hz), 6.09 (d, 1H), 5.99 (d, 1H, J=11.1 Hz), 5.52(dd, 1H, J=8.8 Hz and 15.1 Hz), 5.31 (m, 1H), 4.86 (m, 1H), 4.39 (m,1H), 4.17 (m, 1H), 3.96 (d, 2H), 3.86 (d, OH), 3.68 (t, OH), 3.62 (d,OH), 2.87 (m, 1H), 2.50 (dd, 1H), 2.29 (dd, 1H), 2.20 (m, 1H), 1.72 (bs,3H), 2.10-1.20 (m, 13H), 1.08 (d, 3H), 0.60 (s, 3H) ppm.

Example 91(S),3(R),26-Trihydroxy-9,10-seco-20(S)-cholesta-5(Z),7(E),10(19),22(E),24(E)-penta-ene(Compound 10)

Silyl-protected intermediate: 529 (0.126 g, 0.2 mmol).

Compound 10: ¹³C-NMR: δ=147.7, 143.2, 141.8, 134.5, 132.9, 125.5, 125.0,123.4, 117.0, 111.8, 70.9, 68.7, 66.8, 56.8, 56.2, 46.0, 45.3, 42.9,41.0, 39.8, 29.1, 27.2, 23.5, 22.1, 21.3, 14.2, 12.3 ppm.

Example 101(S),3(R),26-Trihydroxy-9,10-secocholesta-5(Z),7(E),10(19),22(E),24(Z)-penta-ene(Compound 11)

Silyl-protected intermediate: 530 (24 mg, 0.04 mmol).

Compound 11: ¹H-NMR: δ=6.38 (d, 1H), 6.22 (dd, 1H, J=11.1 Hz and 14.9Hz), 6.01 (d, 1H), 5.88 (d, 1H, J=11.1 Hz), 5.51 (dd, 1H, J=8.8 Hz and14.9 Hz), 5.32 (m, 1H), 5.00 (m, 1H), 4.43 (m, 1H), 4.24 (s, 2H), 4.22(m, 1H), 2.83 (m, 1H), 2.60 (dd, 1H), 2.31 (dd, 1H), 2.13 (m, 1H), 1.85(bs, 3H), 1.05 (d, 3H), 2.08-0.80 (m, 16H), 0.56 (s, 3H) ppm.

Example 111(S),3(R),26-Trihydroxy-9,10-seco-20(S)-cholesta-5(Z),7(E),10(19),22(E),24(Z)-penta-ene(Compound 12)

Silyl-protected intermediate: 531 (17 mg, 0.03 mmol).

Compound 12: ¹H-NMR: δ=6.37 (d, 1H), 6.20 (dd, 1H, J=11.1 Hz and 14.9Hz), 6.01 (d, 1H), 5.89 (d, 1H, J=11.1 Hz), 5.52 (dd, 1H, J=9.5 Hz and14.9 Hz), 5.33 (m, 1H), 5.00 (m, 1H), 4.43 (m, 1H), 4.24 (s, 2H), 4.22(m, 1H), 2.81 (m, 1H), 2.60 (dd, 1H), 2.31 (dd, 1H), 1.86 (bs, 3H), 0.94(d, 3H), 2.20-0.80 (m, 17H), 0.50 (s, 3H) ppm.

Example 121(S),3(R)-Dihydroxy-20(R)-(4-methyl-5-ethyl-5-hydroxy-1(E),3(E)-heptadienyl)-9,10-secopregna-5(Z),7(E),10(19)-triene(Compound 13)

Silyl-protected intermediate: 532 (90 mg, 0.13 mmol).

Compound 13: ¹³C-NMR: δ=147.7, 143.0, 140.2, 137.5, 133.0, 125.0, 124.6,124.0, 117.1, 111.8, 78.4, 70.8, 66.8, 56.4, 56.4, 46.0, 45.3, 42.9,40.5, 40.4, 31.8, 29.1, 27.7, 23.6, 22.3, 20.6, 14.2, 13.5, 12.3, 7.7ppm.

Example 131(S),3(R)-Dihydroxy-20(R)-(3-cyclopropyl-1(E),3-butadienyl)-9,10-secopregna-5(Z),7(E),10(19)-triene(Compound 14)

Silyl-protected intermediate: 533 (0.10 mg, 0.16 mmol).

Purification of compound 14 by crystallisation from methyl formate.

Compound 14: ¹³C-NMR: δ=147.8, 147.7, 143.1, 137.0, 133.0, 129.9, 125.0,117.1, 111.8, 110.5, 70.8, 66.9, 56.5, 56.4, 46.0, 45.3, 42.9, 40.4,40.3, 29.1, 27.6, 23.6, 22.2, 20.6, 12.8, 12.3, 5.5 ppm.

Example 141(S),3(R)-Dihydroxy-9,10-secocholesta-5(Z),7(E),10(19),24-tetra-ene-22-yne(Compound 15)

Silyl-protected intermediate: 1002 (100 mg, 0.16 mmol).

Purification of compound 15 by chromatography (50% v:v, ethyl acetate inpetroleum ether).

Compound 15: ¹³C-NMR: δ=147.4, 146.1, 142.6, 132.9, 124.7, 117.0, 111.6,105.4, 96.4, 78.9, 70.6, 66.6, 56.1, 55.8, 45.6, 45.1, 42.7, 39.5, 28.8,28.3, 26.3, 24.4, 23.2, 22.1, 21.6, 20.5, 12.3 ppm.

Example 151(S),3(R)-Dihydroxy-20(R)-(5-methyl-5-hydroxy-1,3-hexadiynyl)-9,10-secopregna-5(Z),7(E),10(19)-triene(Compound 16)

Silyl-protected intermediate: 1004 (41 mg, 0.06 mmol).

Compound 16: ¹³C-NMR: δ=147.7, 142.7, 133.2, 124.9, 117.1, 111.8, 85.8,80.6, 70.8, 67.4, 66.8, 65.6, 65.4, 55.9, 55.6, 45.9, 45.2, 42.9, 39.3,31.2, 29.1, 28.2, 27.3, 23.5, 21.9, 20.5, 12.4 ppm.

Example 161(S),3(R)-Dihydroxy-20(S)-(5-ethyl-5-hydroxy-1,3-heptadiynyl)-9,10-secopregna-5(Z),7(E),10(19)-triene(Compound 17)

Silyl-protected intermediate: 1006 (66 mg, 0.1 mmol)

Compound 17: ¹³C-NMR: δ=147.6, 142.4, 133.4, 124.8, 117.4, 111.9, 85.1,78.8, 72.7, 70.9, 69.4, 66.9, 65.3, 55.9, 55.8, 45.9, 45.3, 42.9, 39.6,34.3, 29.0, 28.4, 26.3, 23.3, 22.2, 21.0, 12.5, 8.5 ppm.

Example 171(S),3(R)-Dihydroxy-20(R)-(5-ethyl-5-hydroxy-1,3-heptadiynyl)-9,10-secopregna-5(Z),7(E),10(19)-triene(Compound 18)

Silyl-protected intermediate: 1008 (69 mg, 0.1 mmol)

Compound 18: ¹³C-NMR: δ=147.7, 142.7, 133.2, 124.9, 117.1, 111.8, 85.2,79.0, 72.7, 70.8, 69.4, 66.9, 65.6, 55.9, 55.6, 45.9, 45.3, 42.9, 39.3,34.3, 29.1, 28.2, 27.3, 23.5, 22.0, 20.4, 12.4, 8.5 ppm.

Example 181(S),3(R)-Dihydroxy-20(S)-(4,4-dibromo-1,3-butadien-1yl)-9,10-seco-pregna-5(Z),7(E),10(19)-triene(Compound 8)

Silyl-protected intermediate: 520 (75 mg, 0.1 mmol)

Purification of compound 8 by chromatography (50% v:v, ethyl acetate inpetroleum ether).

Compound 8: ¹³C-NMR: δ=147.4, 145.7, 142.6, 137.0, 132.9, 124.7, 124.7,116.9, 111.6, 88.1, 70.6, 66.6, 56.4, 55.9, 45.7, 45.0, 42.7, 40.7,39.5, 28.8, 26.9, 23.2, 21.8, 20.5, 12.1 ppm.

Example 191(S)-Fluoro-3(R)-hydroxy-9,10-secocholesta-5(Z),7(E),10(19),22(E),24-penta-ene(Compound 21)

By general procedure 8 (but prolonging the reaction time to 5 h)compound 522 (0.8 g, 2.1 mmol) was desilylated to the correspondingalcohol, 8-beta-hydroxy-desAB-cholesta-22(E),24-diene. A stirredsolution, maintained at about 5° C., of this alcohol (0.39 g, 1.5 mmol)in dry DCM (10 ml) was treated with the Dess-Martin periodinane (0.7 g).After 30 min 5% aq. sodium hydrogen carbonate solution and 10% aq.sodium thiosulphate solution were added and stirring continued for 10min before dilution with ether and performing the standard work-up togive the corresponding ketone, 8-oxo-desAB-cholesta-22(E),24-diene. Thisketone (67 mg, 0.25 mmol) was slowly added in THF (3 ml) to a solution,maintained at about −70° C., of the lithio-derivative (generated in situwith 1 eq. n-BuLi) of phosphine oxide II (A=F, B=CH₂) (0.5 mmol) in dryTHF (4 ml). After stirring at the same temperature for 2 h and warmingto room temperature, the mixture was quenched with wet ether and workedup as standard (chromatography: 2% v:v ether in petroleum ether) to givethe TBS derivative if compound 21. This was desilylated using generalprocedure 9 to give Compound 21.

Compound 21: ¹H-NMR in agreement with structure.

Example 201(S),3(R)-Dihydroxy-19-nor-9,10-secocholesta-5,7(E),22(E),24-tetra-ene(Compound 22)

By general procedure 8 (but prolonging the reaction time to 5 h)compound 522 (0.8 g, 2.1 mmol) was desilylated to the correspondingalcohol, 8-beta-hydroxy-desAB-cholesta-22(E),24-diene. A stirredsolution, maintained at about 5° C., of this alcohol (0.39 g, 1.5 mmol)in dry DCM (10 ml) was treated with the Dess-Martin periodinane (0.7 g).After 30 min 5% aq. sodium hydrogen carbonate solution and 10% aq.sodium thiosulphate solution were added and stirring continued for 10min before dilution with ether and performing the standard work-up togive the corresponding ketone, 8-oxo-desAB-cholesta-22(E),24-diene. Thisketone (67 mg, 0.25 mmol) was slowly added in THF (3 ml) to a solution,maintained at about −70° C., of the lithio-derivative (generated in situwith 1 eq. n-BuLi) of phosphine oxide II (A=O-TBS, B=H₂) (0.5 mmol) indry THF (4 ml). After stirring at the same temperature for 2 h andwarming to room temperature, the mixture was quenched with wet ether andworked up as standard (chromatography: 2% v:v ether in petroleum ether)to give the di-TBS derivative if compound 22. This was desilylated usinggeneral procedure 9 to give Compound 22.

Compound 22: ¹H-NMR in agreement with structure.

Example 211(S),3(S)-Dihydroxy-9,10-secocholesta-5(Z),7(E),10(19),22(E),24-penta-ene(Compound 23)

This compound was prepared analogously to Compound 29, using theintermediate of type 5 that was obtained analogously to Compound 552 bysubstituting acetone for the diethyl ketone.

Compound 23: ¹H-NMR in agreement with structure.

Example 221(S),3(R)-Dihydroxy-9,10-secocholesta-5(Z),7(E),10(19),16,22(E),24-hexa-ene(Compound 24)

This compound was prepared analogously to Compound 1(Example 1), butstarting the sequence (of Preparations 1 and 101) using the intermediateof type 2a that was prepared from 104 analogously to the describedpreparation of 202 from 101 in WO9100855.

Compound 24: ¹H-NMR in agreement with structure.

Example 231(S),3(R)-Dihydroxy-26,26,26,27,27,27-hexafluoro-9,10-secocholesta-5(Z),7(E),10(19),22(E),24-penta-ene(Compound 25)

Silyl-protected intermediate: 535 (55 mg).

Chromatography: 50% v:v ethyl acetate in petroleum ether.

Compound 25: ¹H-NMR (CDCl₃): δ=6.96 (d,1H), 6.43 (t,1H), 6.35 (d,1H),6.18 (dd,1H), 6.01 (d,1H), 5.31 (bs,1H), 4.98 (bs,1H), 4.41 (m,1H), 4.22(m,1H), 2.80 (dd,1H), 2.57 (dd,1H), 2.30 (m,2H), 2.05-1.20 (m,8H), 1.10(d,3H), 0.58 (s,3H) ppm.

Example 24 Capsules containing Compound 1

Compound 1 was dissolved in arachis oil to a final concentration of 10μg of Compound 1/ml oil. 10 Parts by weight of gelatine, 5 parts byweight glycerine, 0.08 parts by weight potassium sorbate, and 14 partsby weight distilled water were mixed together with heating and formedinto soft gelatine capsules. These were then filled each with 100 μl ofCompound 1 in oil solution, such that each capsule contained 1 μg ofCompound 1.

Example 251(S),3(R)-Dihydroxy-20(R)-(4,4-dibromo-1,3-butadien-1-yl)-9,10-seco-pregna-5(Z),7(E),10(19)-triene(Compound 27)

Silyl-protected intermediate: 537 (101 mg, 0.13 mmol).

Purification of compound 27 by chromatography (50% v:v, ethyl acetate inpetroleum ether).

Compound 27: ¹³C-NMR: δ=147.7, 145.3, 142.7, 137.3, 133.2, 125.0, 124.9,117.3, 111.8, 88.3, 70.8, 66.9, 56.2, 55.8, 46.0, 45.3, 42.9, 40.6,40.3, 29.0, 27.6, 23.5, 22.3, 20.0, 12.3 ppm.

Example 261(S),3(R)-Dihydroxy-26,27-dimethyl-9,10-secocholesta-5(Z),7(E),10(19),22(E),24-penta-ene(Compound 28)

Silyl-protected intermediate: 551 (70 mg).

Purification of compound 28 by chromatography: 50% v:v ethyl acetate inpetroleum ether

Compound 28: ¹³C-NMR (CDCl₃): δ=147.4, 143.9, 142.9, 138.4, 132.7,124.8, 123.9, 122.7, 116.9, 111.6, 70.6, 66.7, 56.2, 56.2, 45.7, 45.1,42.7, 40.3, 40.2, 29.3, 28.9, 27.6, 23.6, 23.4, 22.1, 20.6, 13.2, 12.4,12.1 ppm.

Example 271(S),3(S)-Dihydroxy-26,27-dimethyl-9,10-secocholesta-5(Z),7(E),10(19),22(E),24-penta-ene(Compound 29)

Silyl-protected intermediate: 552 (25 mg).

Purification of compound 30 by chromatography: 50% v:v ethyl acetate inpetroleum ether.

Compound 29: ¹H-NMR: δ=6.42 (d,1H), 6.17 (dd,1H), 6.00 (d,1H), 5.70(d,1H), 5.43 (dd,1H), 5.28 (bd,1H), 4.98 (d,1H), 4.29 (m,1H), 4.03(m,1H), 2.84 (dd,1H), 2.64 (bd,1H), 2.55 (dd,1H), 2.43 (dd,1H),2.20-1.15 (m,19H), 1.04 (d,3H), 1.00 (m,6H), 0.56 (s,3H) ppm.

Example 281(S),3(R)-Dihydroxy-24-methyl-26,27-methano-9,10-secocholesta-5(Z),7(E),10(19),22(E),24-penta-ene(Compound 30)

Silyl-protected intermediate: 541 (47 mg, 0.072 mmol).

Purification of compound 30 by chromatography (50% v:v, ethyl acetate inpetroleum ether).

Compound 30: ¹H-NMR: δ=6.38 (d,1H), 6.01 (d,1H), 5.99 (d,1H,J=15.7 Hz),5.33 (m,1H), 5.32 (m,1H), 5.00 (m,1H), 4.43 (m,1H), 4.23 (m,1H), 2.83(dd,1H), 2.79-2.65 (m,4H), 2.60 (dd,1H), 2.31 (dd,1H), 2.22-1.16(m,18H), 1.56 (bs,3H), 1.05 (d,3H), 0.57 (s,3H) ppm.

Example 291(S),3(R)-Dihydroxy-20(R)-(4,4-dichloro-1,3-butadien-1-yl)-9,10-seco-pregna-5(Z),7(E),10(19)-triene(Compound 31)

Silyl-protected intermediate: 539 (70 mg, 0.10 mmol).

Purification of compound 31 by chromatography (50% v:v, ethyl acetate inpetroleum ether).

Compound 31: ¹³C-NMR: δ=147.7, 144.6, 142.6, 133.2, 129.2, 124.9, 122.4,118.9, 117.3, 111.8, 70.8, 66.9, 56.3, 55.9, 46.0, 45.3, 42.9, 40.6,40.3, 29.0, 27.6, 23.5, 22.3, 20.2, 12.3 ppm.

Example 301(S),3(R)-Dihydroxy-26,27-ethano-9,10-secocholesta-5(Z),7(E),10(19),22(E),24-penta-ene(Compound 32)

Silyl-protected intermediate: 543 (300 mg, 0.46 mmol).

Purification of compound 32 by chromatography (50% v:v, ethyl acetate inpetroleum ether).

Compound 32: ¹³C-NMR: δ=147.6, 145.1, 143.1, 137.6, 132.9, 125.7, 125.0,120.4, 117.1, 111.8, 70.8, 66.8, 56.4, 45.9, 45.2, 42.8, 40.5, 40.4,33.9, 29.3, 29.1, 27.8, 26.4, 26.3, 23.6, 22.2, 20.8, 12.3 ppm.

Example 311(S),3(R)-Dihydroxy-26,27-propano-9,10-secocholesta-5(Z),7(E),10(19),22(E),24-penta-ene(Compound 33)

Silyl-protected intermediate: 545 (200 mg, 0.30 mmol).

Purification of compound 22 by chromatography (50% v:v, ethyl acetate inpetroleum ether).

Compound 33: ¹³C-NMR: δ=147.6, 143.1, 140.9, 138.5, 132.9, 125.0, 123.6,122.1, 117.1, 111.8, 70.8, 66.9, 56.5, 56.4, 46.0, 45.3, 42.9, 40.4,37.2, 29.2, 29.1, 28.4, 27.8, 27.7, 26.9, 23.6, 22.3, 20.7, 12.3 ppm.

Example 321(S),3(R)-Dihydroxy-20(S)-cyclopropylidenemethyl-9,10-seco-pregna-5(Z),7(E),10(19)-triene(Compound 34)

Silyl-protected intermediate: 547 (143 mg, 0.24 mmol).

Purification of compound 34 by chromatography (50% v:v, ethyl acetate inpetroleum ether).

Compound 34: ¹³C-NMR: δ=147.7, 143.4, 132.8, 125.1, 125.0, 118.0, 116.9,111.8, 70.9, 66.9, 56.9, 56.1, 46.0, 45.3, 42.9, 39.9, 39.8, 29.2, 27.3,23.6, 22.1, 20.9, 12.4, 2.1, 1.9 mmp.

Example 331(S),3(R)-Dihydroxy-20(R)-cyclopropylidenemethyl-9,10-seco-pregna-5(Z),7(E),10(19)-triene(Compound 35)

Silyl-protected intermediate: 549 (111 mg, 0.18 mmol).

Purification of compound 35 by chromatography (50% v:v, ethyl acetate inpetroleum ether).

Compound 35: ¹³C-NMR: δ=147.7, 143.2, 132.9, 125.0, 124.5, 118.5, 117.0,111.7, 70.8, 66.9, 56.8, 56.4, 45.9, 45.3, 42.9, 40.4, 39.7, 29.1, 27.5,23.6, 22.3, 20.5, 12.2, 2.3, 1.9

Example 3620-Epi-1(S),3(R)-dihydroxy-26,26,26,27,27,27-hexafluoro-9,10-secocholesta-5(Z),7(E),10(19),22(E),24-penta-ene(Compound 36)

This compound was prepared analogously to Compound 25 (Example 23), butstarting the sequence (of Preparations 34 and 128) with20-epi-1(S),3(R)-di-TBS-oxy-9,10-seco-chola-5(E),7(E),10(19),22(E)-tetra-ene-24-ol.

Compound 36: ¹H-NMR in agreement with structure.

The invention claimed is:
 1. A compound according to formula I

in which formula R1 and R2, which are the same, represent halogen,(C₁-C₆)hydrocarbyl, optionally substituted with one or more fluorineatoms, or, together with the carbon atom to which they are bothattached, R1 and R2 form a (C₃-C₆) carbocyclic ring; R3 representshydrogen or methyl; X represents (E)-ethenylene or (Z)-ethenylene; Y andZ independently represent hydrogen or methyl; the bond between C#16 andC#17 is depicted with a dotted line to illustrate that said bond is asingle bond, in which case the projection of the ring substituent isbeta; A represents hydroxyl; B represents CH₂; the configuration in the3-position corresponds to the same configuration as in natural vitaminD₃ (normal), or the configuration in the 3-position is opposite tonatural vitamin D₃ (epi); and in vivo hydrolysable esters and stereoisomeric forms thereof.
 2. A compound according to claim 1 wherein R1and R2 when taken separately, independently represent bromo, chloro,methyl, ethyl, trifluoromethyl, 1-propyl, 2-propyl, cyclopropyl,2-methyl-2-propyl, or 3-pentyl.
 3. A compound according to claim 1wherein R1 and R2 are the same and both represent methyl, ethyl, bromo,or trifluoromethyl.
 4. A compound according to claim 1 wherein R1 and R2when taken together with the carbon atom to which they are both attachedto form a C₃ carbocyclic ring or a C₄ carbocyclic ring.
 5. A compoundaccording to claim 4 wherein R1 and R2 when taken together are ethyleneor tri-methylene, such as R1 and R2 when taken together with the carbonatom to which they are both attached to form a C₃ carbocyclic ring or aC₄ carbocyclic ring.
 6. A compound according to claim 1 wherein R1represents —CMe₃.
 7. A compound according to claim 1 wherein theconfiguration in the 3-position corresponds to the configuration ofnatural vitamin D₃ (normal).
 8. A compound according to claim 1 whereinthe configuration in the 3-position corresponds to the oppositeconfiguration to that of natural vitamin D₃ (epi).
 9. A compoundaccording to claim 1 selected from the list consisting of1(S),3(R)-Dihydroxy-9,10-secocholesta-5(Z),7(E),10(19),22(E),24-penta-ene(Compound 1),1(S),3(R)-Dihydroxy-9,10-secocholesta-5(Z),7(E),10(19),22(Z),24-penta-ene(Compound 2),20(S),1(S),3(R)-Dihydroxy-9,10-secocholesta-5(Z),7(E),10(19),22(E),24-penta-ene(Compound 3),1(S),3(R)-Dihydroxy-9,10-seco-26,27-cyclo-cholesta-5(Z),7(E),10(19),22(E),24-penta-ene(Compound 4),20(S),1(S),3(R)-Dihydroxy-9,10-seco-26,27-cyclo-cholesta-5(Z),7(E),10(19),22(E),24-penta-ene(Compound 5),1(S),3(R)-Dihydroxy-9,10-seco-26,27-methano-cholesta-5(Z),7(E),10(19),22(E),24-penta-ene(Compound 6),20(S),1(S),3(R)-Dihydroxy-9,10-seco-26,27-methano-cholesta-5(Z),7(E),10(19),22(E),24-penta-ene(Compound 7),1(S),3(R)-Dihydroxy-20(S)-(4,4-dibromo-1,3-butadien-1yl)-9,10-seco-pregna-5(Z),7(E),10(19)-triene(Compound 8),1(S),3(S)-Dihydroxy-9,10-secocholesta-5(Z),7(E),10(19)22(E),24-penta-ene(Compound 23),1(S),3(R)-Dihydroxxy-26,26,26,27,27,27-hexafluoro-9,10-secocholesta-5(Z),7(E),10(19),22(E),24-penta-ene (Compound 25),1(S),3(R)-Dihydroxy-20(R)-(4,4-dibromo-1,3-butadien-1-yl)-9,10-seco-pregna-5(Z),7(E),10(19)-triene(Compound 27),1(S),3(R)-Dihydroxy-26,27-dimethyl-9,10-secocholesta-5(Z),7(E),10(19),22(E),24-penta-ene(Compound 28),1(S),3(S)-Dihydroxy-26,27-dimethyl-9,10-secocholesta-5(Z),7(E),10(19),22(E),24-penta-ene(Compound 29),1(S),3(R)-Dihydroxy-24-methyl-26,27-methano-9,10-secocholesta-5(Z),7(E),10(19),22(E),24-penta-ene(Compound 30),1(S),3(R)-Dihydroxy-20(R)-(4,4-dichloro-1,3-butadien-1-yl)-9,10-seco-pregna-5(Z),7(E),10(19)-triene(Compound 31),1(S),3(R)-Dihydroxy-26,27-ethano-9,10-secocholesta-5(Z),7(E),10(19),22(E),24-penta-ene(Compound 32),1(S),3(R)-Dihydroxy-26,27-propano-9,10-secocholesta-5(Z),7(E),10(19),22(E),24-penta-ene(Compound 33), and20(S),1(S),3(R)-Dihydroxy-26,26,26,27,27,27-hexafluoro-9,10-secocholesta-5(Z),7(E),10(19),22(E),24-penta-ene (Compound 36).
 10. A pharmaceutical compositioncomprising a compound according to claim 1, and optionally apharmaceutically acceptable carrier.
 11. A pharmaceutical compositioncomprising a compound according to claim 9, and optionally apharmaceutically acceptable carrier.